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The marine leech Stibarobdella loricata (Harding, 1924) (Hirudinea, Piscicolidae), parasitic on the angel shark Squatina spp. and sandtiger shark Carcharias taurus Rafinesque, 1810 (Chondrichthyes: Squatinidae, Carchariidae) in Southern Brazilian waters.

The presence of the marine leech, Stibarobdella loricata (Harding, 1924) (Hirudinea, Piscicolidae), is reported on the southern coast of Brazil, based on seven lots with 47 specimens, between 71 and 182 mm in total length, collected on the dorsal region of angel sharks, Squatina argentina (Marini, 1930); S. guggenheim Marini, 1936; S. punctata Marini, 1936 (Chondrichthyes, Squatinidae); and on the head of a sandtiger shark, Carcharias taurus Rafinesque, 1810 (Chondrichthyes, Carchariidae). This is the first record of S. loricata in the western Atlantic and of its parasitic association with S. argentina, S. guggenheim, S. punctata, and C. taurus.

Animals↗

Branchial blood flow distribution in the blue shark (Prionace glauca) and the leopard shark (Triakis semifasciata).

Electromagnetic flow (EMF) quantification of total cardiac stroke flow is not feasible for most elasmobranchs because the vascular anatomy precludes probe placement adjacent to the heart and proximal to all afferent branchial arteries (aba). Most previous studies report a fractional cardiac flow, made with the EMF probe placed on the ventral aorta between the innominate arteries and aba 3. Estimation of total cardiac stroke flow from such data requires a flow correction factor obtained by sacrificing the fish, and carrying out a two step in situ/in vitro flow calibration procedure which is based on tenuous assumptions. Ventral aortic blood flow measurements using the EMF techniques were carried out on large blue sharks, and radiographic imaging studies of ventral aortic and branchial blood flow were done on leopard sharks to verify previously estimated fractional cardiac stroke flow correction factors. The innominate flow fraction determined for both species in these studies are similar and agree with previous estimates for elasmobranchs. EMF data for Prionace show 38% of cardiac stroke flow goes to the innominate arteries, 23% into aba 3, 12% into aba 4, and 27% into aba 5. Radiographic analyses with Triakis reveal that 32% of its cardiac stroke volume flows into the innominate arteries which is in agreement with the in situ/in vitro fractional flow estimate (33%).

Animals↗

Partitioning of body fluids in the Lake Nicaragua shark and three marine sharks.

The relative volumes of major body fluids of freshwater and marine sharks are remarkably similar in spite of the differences in external medium and in osmotic pressure of body fluids. The small differences detected are in agreement with differences reported in comparisons of freshwater and marine teleosts: a slightly higher total water content and a smiller ratio of extracellular to intracellular fluids in freshwater forms.

Animals↗

Shark myoglobins I. Isolation and characterization of myoglobins from the sharks, Squalus japonicus and Proscyllium habereri.

Native oxymyoglobins from the sharks, Squalus japonicus and Proscyllium habereri were also isolated directly from red muscle. The essential step was the chromatographic separation of oxymyoglobin from metmyoglobin on a DEAE-cellulose column. The rate of autoxidation of native oxymyoglobin to metmyoglobin was examined over the pH range of 5-12 in 0.1 M buffer at 25 degrees C and the logarithms of the observed first-order rate constants, log (kobs), were plotted as a function of pH. The pH dependence for the autoxidation of Squalus myoglobin showed almost the same profile as those of bovine, sperm whale and yellowfin tuna myoglobins with distal histidines. On the other hand, the pH dependence of Proscyllium myoglobin differed remarkably from those of other myoglobins, especially in the absence of the proton-catalyzed processes in the acidic region of pH. These results suggest that Proscyllium myoglobin lacks distal histidine.

Animals↗

Enzymic sulfation of bile salts. Partial purification and characterization of an enzyme from the liver of the shark Heterodontus portusjacksoni that catalyses the sulfation of the shark bile steroid 5 beta-scymnol.

An enzyme system which catalyses the transfer of the sulfate group from 3'-phosphoadenosine-5'-phosphosulfate to the bile steroid 5 beta-scymnol has been isolated and characterized from the liver of the shark Heterodontus portusjacksoni (Meyer 1793). The enzyme is present in the cytosol fraction of liver cells. It was partially purified by hydroxylapatite chromatography, molecular sizing by G100-Sephadex and isoelectrofocusing electrophoresis. The apparent Km value for 3'-phosphoadenosine-5'-phosphosulfate was 4 microM and that for 5 beta-scymnol, 14 microM. The enzyme activity is inhibited by iodoacetate and p-chloromercuribenzoate indicating the possible requirement of a sulfhydryl group for activity. The molecular weight of the enzyme was estimated to be 40 kDa by gel filtration. This was verified by running the partially purified material on a native gel and electrophoretically separating two major bands corresponding to molecular weights of 40 and 45 kDa, respectively. Isoelectric focusing of the purified material resulted in two major bands with pI values of 5.0 and 5.85. Enzymatic activity was found to be optimal at a pH of 6.5 with little activity recorded at pH 5.0 and 8.0.

Animals↗

Morphology and evolution of the jaw suspension in lamniform sharks.

The morphology of the jaw suspension and jaw protrusion mechanism in lamniform sharks is described and mapped onto a cladogram to investigate how changes in jaw suspension and protrusion have evolved. This has revealed that several evolutionary modifications in the musculoskeletal apparatus of the jaws have taken place among lamniform sharks. Galeomorph sharks (Carcharhiniformes, Lamniformes, Orectolobiformes, and Heterodontiformes) have paired ethmopalatine ligaments connecting the ethmoid process of the upper jaw to the ethmoid region of the cranium. Basal lamniform sharks also acquired a novel single palatonasal ligament connecting the symphysis of the upper jaw to the cranium mid-ventral to the nasal capsule. Sharks in the family Lamnidae subsequently lost the original paired ethmopalatine ligament while retaining the novel palatonasal ligament. Thus, basal lamniform taxa (Mitsukurina owstoni, Carcharius taurus, Alopias vulpinnis) have increased ligamentous support of the lateral region of the upper jaw while derived species (Lamnidae) have lost this lateral support but gained anterior support. In previous studies the morphology of the jaw suspension has been shown to play a major role in the mechanism of upper jaw protrusion in elasmobranchs. The preorbitalis is the primary muscle effecting upper jaw protrusion in squalean (sister group to galeomorphs) and carcharhiniform (sister group to lamniforms) sharks. The preorbitalis originates from the quadratomandibularis muscle and inserts onto the nasal capsule in squalean and carcharhiniform sharks. Carcharhiniform sharks have evolved a subdivided preorbitalis muscle with the new division inserting near the ethmoid process of the palatoquadrate (upper jaw). Alopid sharks have also independently evolved a partially subdivided preorbitalis with the new division inserting at the base of the ethmoid process and surrounding connective tissue. Lamnid sharks have retained the two preorbitalis divisions but have modified both of the insertion points. The original ventral preorbitalis division now inserts onto the connective tissue surrounding the mid-region of the upper jaw, while the new dorsal preorbitalis division inserts onto the surrounding connective tissue and skin at a more posterior position on the upper jaw. The retractor muscle of the jaws, the levator hyomandibularis, has also been modified during the evolution of lamniform sharks. In most sharks, including basal lamniforms, the levator hyomandibularis inserts onto the hyomandibula and functions to retract the jaws after protrusion. In alopid and lamnid sharks the levator hyomandibularis inserts primarily onto the upper and lower jaws around the jaw joint and is a more direct route for retracting the jaws. Thus, there has been at least one instance of character loss (ethmopalatine ligament), acquisition (palatonasal ligament), subdivision (preorbitalis), and modification (ventral preorbitalis, dorsal preorbitalis, and levator hyomandibularis) in the ligaments and muscles associated with the jaw suspension and jaw protrusion mechanism in lamniform sharks. While derived lamniform sharks (Lamna nasus, Carcharodon carcharius, and Isurus oxyrinchus) lost the ancestral passive lateral support of the ethmoid articulation of the upper jaw, they simultaneously acquired muscular support by way of the levator hyomandibularis, which provides a dynamic mechanism for lateral support. The evolution of multiple divisions of preorbitalis insertions onto the palatoquadrate and modification of the levator hyomandibularis insertion directly onto the jaws provides an active mechanism for multiple protractions and retractions of the upper jaw, which is advantageous in those sharks that gouge or saw pieces from large oversized prey items.

Animals↗

Function of the heterocercal tail in sharks: quantitative wake dynamics during steady horizontal swimming and vertical maneuvering.

The function of the heterocercal tail in sharks has long been debated in the literature. Previous kinematic data have supported the classical theory which proposes that the beating of the heterocercal caudal fin during steady horizontal locomotion pushes posteroventrally on the water, generating a reactive force directed anterodorsally and causing rotation around the center of mass. An alternative model suggests that the heterocercal shark tail functions to direct reaction forces through the center of mass. In this paper, we quantify the function of the tail in two species of shark and compare shark tail function with previous hydrodynamic data on the heterocercal tail of sturgeon Acipenser transmontanus. To address the two models of shark heterocercal tail function, we applied the technique of digital particle image velocimetry (DPIV) to quantify the wake of two species of shark swimming in a flow tank. Both steady horizontal locomotion and vertical maneuvering were analyzed. We used DPIV with both horizontal and vertical light sheet orientations to quantify patterns of wake velocity and vorticity behind the heterocercal tail of leopard sharks (Triakis semifasciata) and bamboo sharks (Chiloscyllium punctatum) swimming at 1.0Ls(-1), where L is total body length. Two synchronized high-speed video cameras allowed simultaneous measurement of shark body position and wake structure. We measured the orientation of tail vortices shed into the wake and the orientation of the central jet through the core of these vortices relative to body orientation. Analysis of flow geometry indicates that the tail of both leopard and bamboo shark generates strongly tilted vortex rings with a mean jet angle of approximately 30 degrees below horizontal during steady horizontal swimming. The corresponding angle of the reaction force is much greater than body angle (mean 11 degrees ) and the angle of the path of motion of the center of mass (mean approximately 0 degrees ), thus strongly supporting the classical model of heterocercal tail function for steady horizontal locomotion. Vortex jet angle varies significantly with body angle changes during vertical maneuvering, but sharks show no evidence of active reorientation of jet angle relative to body angle, as was seen in a previous study on the function of sturgeon tail. Vortex jet orientation is significantly more inclined than the relatively horizontal jet generated by sturgeon tail vortex rings, demonstrating substantial differences in function in the heterocercal tails of sharks and sturgeon. We present a summary of forces on a swimming shark integrating data obtained here on the tail with previous data on pectoral fin and body function. Body orientation plays a critical role in the overall force balance and compensates for torques generated by the tail. The pectoral fins do not generate lift during steady horizontal locomotion, but play an important hydrodynamic role during vertical maneuvering.

Animals↗

Comparative studies of high performance swimming in sharks I. Red muscle morphometrics, vascularization and ultrastructure.

Tunas (family Scombridae) and sharks in the family Lamnidae are highly convergent for features commonly related to efficient and high-performance (i.e. sustained, aerobic) swimming. High-performance swimming by fishes requires adaptations augmenting the delivery, transfer and utilization of O(2) by the red myotomal muscle (RM), which powers continuous swimming. Tuna swimming performance is enhanced by a unique anterior and centrally positioned RM (i.e. closer to the vertebral column) and by structural features (relatively small fiber diameter, high capillary density and greater myoglobin concentration) increasing O(2) flux from RM capillaries to the mitochondria. A study of the structural and biochemical features of the mako shark (Isurus oxyrinchus) RM was undertaken to enable performance-capacity comparisons of tuna and lamnid RM. Similar to tunas, mako RM is positioned centrally and more anterior in the body. Another lamnid, the salmon shark (Lamna ditropis), also has this RM distribution, as does the closely related common thresher shark (Alopias vulpinus; family Alopiidae). However, in both the leopard shark (Triakis semifasciata) and the blue shark (Prionace glauca), RM occupies the position where it is typically found in most fishes; more posterior and along the lateral edge of the body. Comparisons among sharks in this study revealed no differences in the total RM quantity (approximately 2-3% of body mass) and, irrespective of position within the body, RM scaling is isometric in all species. Sharks thus have less RM than do tunas (4-13% of body mass). Relative to published data on other shark species, mako RM appears to have a higher capillary density, a greater capillary-to-fiber ratio and a higher myoglobin concentration. However, mako RM fiber size does not differ from that reported for other shark species and the total volume of mitochondria in mako RM is similar to that reported for other sharks and for tunas. Lamnid RM properties thus suggest a higher O(2) flux capacity than in other sharks; however, lamnid RM aerobic capacity appears to be less than that of tuna RM.

Adaptation, Biological↗

Evolution of high-performance swimming in sharks: transformations of the musculotendinous system from subcarangiform to thunniform swimmers.

In contrast to all other sharks, lamnid sharks perform a specialized fast and continuous "thunniform" type of locomotion, more similar to that of tunas than to any other known shark or bony fish. Within sharks, it has evolved from a subcarangiform mode. Experimental data show that the two swimming modes in sharks differ remarkably in kinematic patterns as well as in muscle activation patterns, but the morphology of the underlying musculotendinous system (red muscles and myosepta) that drives continuous locomotion remains largely unknown. The goal of this study was to identify differences in the musculotendinous system of the two swimming types and to evaluate these differences in an evolutionary context. Three subcarangiform sharks (the velvet belly lantern shark, Etmopterus spinax, the smallspotted catshark, Scyliorhinus canicula, and the blackmouth catshark, Galeus melanostomus) from the two major clades (two galeans, one squalean) and one lamnid shark, the shortfin mako, Isurus oxyrhinchus, were compared with respect to 1) the 3D shape of myomeres and myosepta of different body positions; 2) the tendinous architecture (collagenous fiber pathways) of myosepta from different body positions; and 3) the association of red muscles with myoseptal tendons. Results show that the three subcarangiform sharks are morphologically similar but differ remarkably from the lamnid condition. Moreover, the "subcarangiform" morphology is similar to the condition known from teleostomes. Thus, major features of the "subcarangiform" condition in sharks have evolved early in gnathostome history: Myosepta have one main anterior-pointing cone and two posterior-pointing cones that project into the musculature. Within a single myoseptum cones are connected by longitudinally oriented tendons (the hypaxial and epaxial lateral and myorhabdoid tendons). Mediolaterally oriented tendons (epineural and epipleural tendons; mediolateral fibers) connect vertebral axis and skin. An individual lateral tendon spans only a short distance along the body (a fraction between 0.05 and 0.075 of total length, L, of the shark). This span is similar in all tendons along the body. Red muscles insert into the midregion of the lateral tendons. The shortfin mako differs substantially from this condition in several respects: Red muscles are internalized and separated from white muscles by a sheath of lubricative connective tissue. They insert into the anterior part of the hypaxial lateral tendon. Rostrocaudally, this tendon becomes very distinct and its span increases threefold (0.06L anteriorly to 0.19L posteriorly). Mediolateral fibers do not form distinct epineural/epipleural tendons in the mako. Since our morphological findings are in good accordance with experimental data it seems likely that the thunniform swimming mode has evolved along with the described morphological specializations.

Animals↗

Concentrations and hazard assessment of polychlorinated biphenyls and organochlorine pesticides in shark liver from the Mediterranean Sea.

Concentrations of polychlorinated biphenyls (PCBs) and organochlorine pesticides (DDTs), were determined in the liver of two different shark species Prionace glauca (blue shark) and Dalatias licha (kitefin shark) from the Mediterranean Sea. In blue shark liver, the concentrations of PCBs (2482 ngg(-1)) and DDTs (2392 ngg(-1)) were comparable, while in kitefin shark the hepatic concentrations of DDTs (4554 ngg(-1)) were significantly higher than those of PCBs (1827 ngg(-1)). Contamination levels differed between species, with kitefin shark showing consistently higher concentrations of DDTs, and blue shark higher levels of PCBs. Congener-specific PCB profiles, similar between the two species were dominated by the higher chlorinated congeners (hexachlorobiphenyls: 62.8-63.9%, penta-: 15.2-21.3%, hepta-: 13.4-14.5%) with most of the lower chlorinated congeners being absent or present at very low levels. In both species, the total 2,3,7,8-tetrachlorodibenz-p-dioxin toxic equivalent (TEQs) concentrations (blue shark: 2.51 pg/g; kitefin shark: 1.46 pg/g) seem to be relatively modest. Regards to DDT component pattern, p,p'-DDE was dominant in the liver of both species (blue shark: 81.5%; kitefin shark: 38.0%), while the percentage composition of the other metabolites was differently characterized. The composition pattern of DDTs and the low value of p,p'-DDE/DDTs ratio in the specimens from Ionian Sea suggest that organochlorine pesticide contamination is still continuing in this marine environment.

Animals↗

Three-dimensional kinematics and wake structure of the pectoral fins during locomotion in leopard sharks Triakis semifasciata.

The classical theory of locomotion in sharks proposes that shark pectoral fins are oriented to generate lift forces that balance the moment produced by the oscillating heterocercal tail. Accordingly, previous studies of shark locomotion have used fixed-wing aircraft as a model assuming that sharks have similar stability and control mechanisms. However, unlike airplanes, sharks are propelled by undulations of the body and tail and have considerable control of pectoral fin motion. In this paper, we use a new approach to examine the function of the pectoral fins of leopard sharks, Triakis semifasciata, during steady horizontal swimming at speeds of 0.5-2.0ls(-1), where l is total body length, and during vertical maneuvering (rising and sinking) in the water column. The planar orientation of the pectoral fin was measured using three-dimensional kinematics, while fluid flow in the wake of the pectoral fin and forces exerted on the water by the fin were quantified using digital particle image velocimetry (DPIV). Steady horizontal swimming in leopard sharks is characterized by continuous undulations of the body with a positive body tilt to the flow that decreases from a mean of 11 degrees to 0.6 degrees with increasing flow speeds from 0. 5 to 2.0ls(-1). Three-dimensional analysis showed that, during steady horizontal locomotion, the pectoral fins are cambered, concave downwards, at a negative angle of attack that we predict to generate no significant lift. Leopard shark pectoral fins are also oriented at a substantial negative dihedral angle that amplifies roll moments and hence promotes rapid changes in body position. Vortices shed from the trailing edge of the pectoral fin were detected only during vertical maneuvering. Starting vortices are produced when the posterior plane of the pectoral fin is actively flipped upwards or downwards to initiate rising or sinking, respectively, in the water column. The starting vortex produced by the pectoral fin induces a pitching moment that reorients the body relative to the flow. Body and pectoral fin surface angle are altered significantly when leopard sharks change vertical position in the water column. Thus, locomotion in leopard sharks is not analogous to flight in fixed-wing aircraft. Instead, a new force balance for swimming leopard sharks is proposed for steady swimming and maneuvering. Total force balance on the body is adjusted by altering the body angle during steady swimming as well as during vertical maneuvering, while the pectoral fins appear to be critical for initiating maneuvering behaviors, but not for lift production during steady horizontal locomotion.

Animals↗

Differences in hematocrit of blood samples obtained from two venipuncture sites in sharks.

OBJECTIVE: To evaluate differences in Hct between 2 venipuncture sites in captive and free-ranging sharks. ANIMALS: 32 healthy adult captive sharks (Carcharhinus melanopterus, Carcharhinus plumbeus, Stegastoma fasciatum, Orectolobus japonicus, and Triaenodon obesus) and 15 captured free-ranging adult sharks (Carcharhinus limbatus and Carcharhinus acronotus). PROCEDURES: Blood samples were collected from the caudal tail artery followed by collection from the sinus located immediately caudal to the cranial dorsal fin. The Hct was determined for each sample and results were compared. Additionally, results for sharks that were highly active and used aerobic metabolism were compared with results for sharks that were less active and tolerant of anaerobic conditions. RESULTS: Mean Hct for all sharks was significantly less (8% less) in blood samples obtained from the cranial dorsal fin sinus, compared with the Hct for samples obtained from the caudal tail artery. When compared on the basis of metabolic class, sharks that were more tolerant of anaerobic conditions had lower Hct values and smaller differences between the 2 venipuncture sites. CONCLUSIONS AND CLINICAL RELEVANCE: Hct values were significantly lower in blood samples collected from the cranial dorsal fin sinus compared with values for samples collected from the caudal tail artery. It is important to recognize this difference when evaluating hematologic variables in sharks and when establishing reference ranges for Hcts for shark populations. Sharks that were more active and relied on aerobic metabolism had higher Hct values than did anaerobic-tolerant sharks, and the difference in Hct values between venipuncture sites was more pronounced.

Analysis of Variance↗

Kinematics and critical swimming speed of juvenile scalloped hammerhead sharks

Kinematics and critical swimming speed (Ucrit) of juvenile scalloped hammerhead sharks Sphyrna lewini were measured in a Brett-type flume (635 l). Kinematic parameters were also measured in sharks swimming in a large pond for comparison with those of sharks swimming in the flume. Sharks in the flume exhibited a mean Ucrit of 65±11 cm s-1 (± s.d.) or 1.17±0.21 body lengths per second (L s-1), which are similar to values for other species of sharks. In both the flume and pond, tailbeat frequency (TBF) and stride length (LS) increased linearly with increases in relative swimming speed (Urel=body lengths traveled per second). In the flume, tailbeat amplitude (TBA) decreased with increasing speed whereas TBA did not change with speed in the pond. Differences in TBF and LS between sharks swimming in the flume and the pond decreased with increases in Urel. Sharks swimming at slow speeds (e.g. 0.55 L s-1) in the pond had LS 19 % longer and TBF 21 % lower than sharks in the flume at the same Urel. This implies that sharks in the flume expended more energy while swimming at comparable velocities. Comparative measurements of swimming kinematics from sharks in the pond can be used to correct for effects of the flume on shark swimming kinematics and energetics.

Journal Article↗

Evaluation of shark cartilage in patients with advanced cancer: a North Central Cancer Treatment Group trial.

BACKGROUND: Shark cartilage has been a popular complementary or alternative medicine intervention. The basis for this popularity is the claim that sharks rarely get cancer because of the high proportion of cartilage in the shark's body. However, early studies were equivocal. Therefore, a clinical trial was conducted to look at the impact of shark cartilage in patients with advanced cancer. The primary goal of this trial was to determine whether a shark cartilage product improved overall survival for patients with advanced cancer who were getting standard care. Secondary research goals were to evaluate toxicities, tolerability, and quality of life associated with this shark cartilage product. METHODS: The study was a two-arm, randomized, placebo-controlled, double-blind, clinical trial. Patients with incurable breast or colorectal carcinoma had to have good performance status and organ function. Patients could be receiving chemotherapy. Patients were all to receive standard care and then to be randomly selected to receive either a shark cartilage product or an identical-appearing and smelling placebo 3 to 4 times each day. RESULTS: Data on a total of 83 evaluable patients were analyzed. There was no difference in overall survival between patients receiving standard care plus a shark cartilage product versus standard care plus placebo. Likewise, there was no suggestion of improvement in quality of life for patients receiving the shark cartilage, compared with those receiving placebo. CONCLUSION: This trial was unable to demonstrate any suggestion of efficacy for this shark cartilage product in patients with advanced cancer.

Aged↗

Characterization of gamma S-crystallin isoforms from lip shark (Chiloscyllium colax): evolutionary comparison between gamma S and beta/gamma crystallins.

gamma S-Crystallin from shark eye lenses, formerly termed beta s crystallin in mammalian lenses, is structurally characterized in this study by cDNA cloning and sequencing. To facilitate sequence characterization of gamma S-crystallin possessing intermediate structural properties between beta- and gamma-crystallins, cDNA mixture was constructed from the poly(A)+ mRNA isolated from shark eye lenses, and amplification by polymerase chain reaction (PCR) was carried out to obtain nucleotide segments encoding multiple shark gamma S-crystallins. Sequencing several positive clones revealed that a multiplicity of isoforms exists in the gamma S-crystallin class of this cartilaginous fish, similar to authentic gamma-crystallin family characterized from the same shark species. Comparison of protein sequences encoded by two representative shark gamma S1 and gamma S2 cDNAs with those published sequences of beta-, gamma-, and gamma S crystallins from bovine, human, bullfrog and carp lenses indicated that there is about 35-64% sequence homology between shark gamma S crystallins and structurally related crystallins from different evolutionary classes, with a higher sequence similarity between shark gamma S and mammalian gamma-crystallins than that of shark gamma S and carp gamma S or bovine gamma S crystallins. A phylogenetic tree constructed on the basis of the sequence divergence among various beta-, gamma-, and gamma S crystallins corroborates the closer relatedness of shark gamma S to authentic gamma-crystallin than to mammalian and teleostean gamma S crystallins. It further strengthens the supposition that ancestral precursors of gamma S-crystallins were present in the shark lens long before the appearance of present-day teleostean and mammalian gamma S-crystallins.

Amino Acid Sequence↗

Functional significance of the shark Na,K-ATPase N-terminal domain. Is the structurally variable N-Terminus involved in tissue-specific regulation by FXYD proteins?

The proteolytic profile after mild controlled trypsin cleavage of shark rectal gland Na,K-ATPase was characterized and compared to that of pig kidney Na,K-ATPase, and conditions for achieving N-terminal cleavage of the alpha-subunit at the T(2) trypsin cleavage site were established. Using such conditions, the shark enzyme N-terminus was much more susceptible to proteolysis than the pig enzyme. Nevertheless, the maximum hydrolytic activity was almost unaffected for the shark enzyme, whereas it was significantly decreased for the pig kidney enzyme. The apparent ATP affinity was unchanged for shark but increased for pig enzyme after N-terminal truncation. The main common effect following N-terminal truncation of shark and pig Na,K-ATPase is a shift in the E(1)-E(2) conformational equilibrium toward E(1). The phosphorylation and the main rate-limiting E(2) --> E(1) step are both accelerated after N-terminal truncation of the shark enzyme, but decreased significantly in the pig kidney enzyme. Some of the kinetic differences, like the acceleration of the phosphorylation reaction, following N-terminal truncation of the two preparations may be due to the fact that under the conditions used for N-terminal truncation, the C-terminal domain of the FXYD regulatory protein of the shark enzyme, PLMS or FXYD10, was also cleaved, whereas the gamma or FXYD2 of the pig enzyme was not. In the shark enzyme, N-terminal truncation of the alpha-subunit abolished association of exogenous PLMS with the alpha-subunit and the functional interactions were abrogated. Moreover, PKC phosphorylation of the preparation, which relieves PLMS inhibition of Na,K-ATPase activity, exposed the N-terminal trypsin cleavage site. It is suggested that PLMS interacts functionally with the N-terminus of the shark Na,K-ATPase to control the E(1)-E(2) conformational transition of the enzyme and that such interactions may be controlled by regulatory protein kinase phosphorylation of the N-terminus. Such interactions are likely in shark enzyme where PLMS has been demonstrated by cross-linking to associate with the Na,K-ATPase A-domain.

Amino Acid Sequence↗

Shark complement: an assessment.

The classical (CCP) and alternative (ACP) pathways of complement activation have been established for the nurse shark (Ginglymostoma cirratum). The isolation of a cDNA clone encoding a mannan-binding protein-associated serine protease (MASP)-1-like protein from the Japanese dogfish (Triakis scyllia) suggests the presence of a lectin pathway. The CCP consists of six functionally distinct components: C1n, C2n, C3n, C4n, C8n and C9n, and is activated by immune complexes in the presence of Ca++ and Mg++ ions. The ACP is antibody independent, requiring Mg++ ions and a heat-labile 90 kDa factor B-like protein for activity. Proteins considered homologues of C1q, C3 and C4 (C2n) of the mammalian complement system have been isolated from nurse shark serum. Shark C1q is composed of at least two chain types each showing 50% identity to human C1q chains A and B. Partial sequence of the globular domain of one of the chains shows it to be C1q-like rather than like mannan-binding protein. N-terminal amino acid sequences of the alpha and beta chain of shark C3 and C4 molecules show significant identity with corresponding human C3 and C4 chains. A sequence representing shark C4 gamma chain, shows little similarity to human C4 gamma chain. The terminal shark components C8n and C9n are functional analogues of mammalian C8 and C9. Anaphylatoxin activity has been demonstrated in activated shark serum, and porcine C5a desArg induces shark leucocyte chemotaxis. The deduced amino acid sequence of a partial C3 cDNA clone from the nurse shark shows 50%, 30% and 24% homology with the corresponding region of mammalian C3, C4 and alpha 2-macroglobulin. Deduced amino acid sequence data from partial Bf/C2 cDNA clones, two from the nurse shark and one from the Japanese dogfish, suggest that at least one species of elasmobranch has two distinct Bf/C2 genes.

Amino Acid Sequence↗