Biomedical subjects

J H Waite

Publications and source records attributed to J H Waite.

At least 19 recordsLinked to original sources

PubMed2006

Role of melanin in mechanical properties of Glycera jaws.

The remarkable mechanical prowess of the jaws of the bloodworm Glycera dibranchiata appears to be a consequence of a robust cross-linked network of organic molecules, notably melanin and proteins, as well as small amounts of unmineralized Cu and a Cu-based mineral. The present study focuses on the role of melanin. Mechanical properties of untreated jaws and the constituent melanin are probed through nanoindentation, both in air and underwater. Complementary information is obtained from density and porosity measurements and attempts at Cu removal from the jaws using EDTA, an effective metal chelator in most biological systems. In near-tip regions of the jaws, mechanical properties attain the highest values and diminish only slightly when wet (by 15-25%), in contrast to the behavior of other organic biomaterials. The melanin constituent contributes significantly to the mechanical integrity of the jaw; its hardness and elastic modulus are about half those of untreated jaws. Although melanin may be the dominant shape-determining component of the structure, it remains to be shown whether jaw assembly is mediated by protein deposition on a melanin scaffold or, conversely, by melanin deposition on a protein scaffold. The inability of EDTA to chelate Cu from the jaws and the relatively high density of the jaws and the melanin support the notion of a highly cross-linked molecular structure. Finally, based on the metric H(3)/E(2) (H being hardness and E the Young's modulus), the results suggest that the abrasion resistance of the jaws is superior to all engineering polymers and competitive with the hardest metallic alloys.

Animals↗

PubMed2005

Oxygen ions observed near Saturn's A ring.

Ions were detected in the vicinity of Saturn's A ring by the Ion and Neutral Mass Spectrometer (INMS) instrument onboard the Cassini Orbiter during the spacecraft's passage over the rings. The INMS saw signatures of molecular and atomic oxygen ions and of protons, thus demonstrating the existence of an ionosphere associated with the A ring. A likely explanation for these ions is photoionization by solar ultraviolet radiation of neutral O2 molecules associated with a tenuous ring atmosphere. INMS neutral measurements made during the ring encounter are dominated by a background signal.

Atmosphere↗

PubMed2002

A pulsating auroral X-ray hot spot on Jupiter.

Jupiter's X-ray aurora has been thought to be excited by energetic sulphur and oxygen ions precipitating from the inner magnetosphere into the planet's polar regions. Here we report high-spatial-resolution observations that demonstrate that most of Jupiter's northern auroral X-rays come from a 'hot spot' located significantly poleward of the latitudes connected to the inner magnetosphere. The hot spot seems to be fixed in magnetic latitude and longitude and occurs in a region where anomalous infrared and ultraviolet emissions have also been observed. We infer from the data that the particles that excite the aurora originate in the outer magnetosphere. The hot spot X-rays pulsate with an approximately 45-min period, a period similar to that reported for high-latitude radio and energetic electron bursts observed by near-Jupiter spacecraft. These results invalidate the idea that jovian auroral X-ray emissions are mainly excited by steady precipitation of energetic heavy ions from the inner magnetosphere. Instead, the X-rays seem to result from currently unexplained processes in the outer magnetosphere that produce highly localized and highly variable emissions over an extremely wide range of wavelengths.

Journal Article↗

PubMed2002

Transient aurora on Jupiter from injections of magnetospheric electrons.

Energetic electrons and ions that are trapped in Earth's magnetosphere can suddenly be accelerated towards the planet. Some dynamic features of Earth's aurora (the northern and southern lights) are created by the fraction of these injected particles that travels along magnetic field lines and hits the upper atmosphere. Jupiter's aurora appears similar to Earth's in some respects; both appear as large ovals circling the poles and both show transient events. But the magnetospheres of Jupiter and Earth are so different---particularly in the way they are powered---that it is not known whether the magnetospheric drivers of Earth's aurora also cause them on Jupiter. Here we show a direct relationship between Earth-like injections of electrons in Jupiter's magnetosphere and a transient auroral feature in Jupiter's polar region. This relationship is remarkably similar to what happens at Earth, and therefore suggests that despite the large differences between planetary magnetospheres, some processes that generate aurorae are the same throughout the Solar System.

Journal Article↗

PubMed2002

Ultraviolet emissions from the magnetic footprints of Io, Ganymede and Europa on Jupiter.

Io leaves a magnetic footprint on Jupiter's upper atmosphere that appears as a spot of ultraviolet emission that remains fixed underneath Io as Jupiter rotates. The specific physical mechanisms responsible for generating those emissions are not well understood, but in general the spot seems to arise because of an electromagnetic interaction between Jupiter's magnetic field and the plasma surrounding Io, driving currents of around 1 million amperes down through Jupiter's ionosphere. The other galilean satellites may also leave footprints, and the presence or absence of such footprints should illuminate the underlying physical mechanism by revealing the strengths of the currents linking the satellites to Jupiter. Here we report persistent, faint, far-ultraviolet emission from the jovian footprints of Ganymede and Europa. We also show that Io's magnetic footprint extends well beyond the immediate vicinity of Io's flux-tube interaction with Jupiter, and much farther than predicted theoretically; the emission persists for several hours downstream. We infer from these data that Ganymede and Europa have persistent interactions with Jupiter's magnetic field despite their thin atmospheres.

Journal Article↗

PubMed2001

An auroral flare at Jupiter.

Jupiter's aurora is the most powerful in the Solar System. It is powered largely by energy extracted from planetary rotation, although there seems also to be a contribution from the solar wind. This contrasts with Earth's aurora, which is generated through the interaction of the solar wind with the magnetosphere. The major features of Jupiter's aurora (based on far-ultraviolet, near-infrared and visible-wavelength observations) include a main oval that generally corotates with the planet and a region of patchy, diffuse emission inside the oval on Jupiter's dusk side. Here we report the discovery of a rapidly evolving, very bright and localized emission poleward of the northern main oval, in a region connected magnetically to Jupiter's outer magnetosphere. The intensity of the emission increased by a factor of 30 within 70 s, and then decreased on a similar timescale, all captured during a single four-minute exposure. This type of flaring emission has not previously been reported for Jupiter (similar, but smaller, transient events have been observed at Earth), and it may be related directly to changes in the solar wind.

Journal Article↗

PubMed2001

Polyphosphoprotein from the adhesive pads of Mytilus edulis.

Achieving a satisfactory biochemical explanation for the opportunistic underwater adhesion of marine invertebrates such as mussels and barnacles requires a detailed characterization of proteins extracted from holdfast structures produced by these organisms. Mefp-5 is an adhesive protein derived from the foot of the common mussel, Mytilus edulis, and deposited into the byssal attachment pads. Purification and primary structure of mefp-5 was determined by peptide mapping and cDNA sequencing. The protein is 74 residues long and has a mass of about 9500 Da. Mefp-5 composition shows a strong amino acid bias: aromatic amino acids, lysine, and glycine represent 65 mol % of the composition. More than a third of all the residues in the protein are posttranslationally modified by hydroxylation or phosphorylation. The conversion of tyrosine to 3, 4-dihydroxyphenyl-L-alanine (DOPA) and serine to O-phosphoserine accounts for the hydroxylation and phosphorylation, respectively. Neither modification is complete since variations in the extent of phosphorylation and hydroxylation can be detected by mass spectrometry. More than 75% of the DOPA is adjacent to basic residues, e.g., Lys-DOPA and DOPA-Lys. Phosphoserine occurs in sequences strikingly reminiscent of acidic mineral-binding motifs that appear in statherin, osteopontin, and others. This may be an adaptation for adhesion to the most common substrata for mussels, i.e., calcareous materials.

Amino Acid Sequence↗

PubMed2001

Oxidative stress and the mechanical properties of naturally occurring chimeric collagen-containing fibers.

The byssal threads of marine mussels are a fiber-reinforced composite material. Fibers are continuous, separated by matrix, and consist of chimeric collagens that encompass within the same primary protein structure domains corresponding to collagen, polyhistidine, and either elastin or dragline spider silk. The elastic modulus (stiffness) of the proximal portion of byssal threads was measured by cyclic stress-strain analysis at 50% extension. Before measurement, the threads were conditioned by various treatments, particularly agitation in aerated or nitrogen-sparged seawater. Stiffness can be permanently increased by more than two times, e.g., from 25 MPa to a maximum of 65 MPa, by simple agitation in aerated seawater. Much but not all of this stiffening can be prevented by agitation under nitrogen. Reversible strain stiffening would seem to be a useful adaptation to lower residual stresses arising from the deformation of two joined materials, i.e., distal and proximal portions with rather different elastic moduli. The permanent strain stiffening that characterizes proximal byssal threads subjected to oxidative stress is probably due to protein cross-linking. In the short term, this results in a stronger thread but at the expense of dynamic interactions between the molecules in the structure.

Animals↗

PubMed2001

Development of the vitellaria of the liver fluke, Fasciola hepatica in the rat host.

The development of the vitellaria of Fasciola hepatica within the liver of its rat host was studied by means of whole-mount stained preparations and transmission electron microscopy, together with light and electron immunocytochemistry using an antibody to vitelline protein B, an eggshell precursor protein synthesized by F. hepatica. No vitelline cells could be identified in flukes recovered from the liver parenchyma, by any of the methods used. In contrast, follicles were present in flukes at the earliest time of recovery from the bile duct, namely, 5 weeks 3 days post-infection. The vitellaria in these flukes formed a row of small follicles on either side of the body. Development of the follicles was rapid: by 6 weeks 3 days, the vitellaria resembled those in the adult fluke and eggs were present in the uterus. Immunolabelling was confined to the shell protein globules in the vitelline cells, confirming the packaging of the eggshell protein within the shell globule clusters.

Animals↗

PubMed2001

Yield and post-yield behavior of mussel byssal thread: a self-healing biomolecular material.

Byssal threads provide marine mussels with the tenacity to remain sessile in habitats of high flow. Under uniaxial tension, byssal threads are typical of other biological and synthetic fibers in exhibiting an initial linear region followed by yield. They differ, however, in their capacity to recover or "self-heal" following yield. We have examined the effect of urea, dithiothreitol (DTT), and metal-chelating agents such as ethylenediamine tetraacetic acid (EDTA) in perturbing the modulus, yield point, and energy dissipated in distal byssal threads stretched cyclically in seawater to a strain of 0.7. Threads stretched in the presence of 8 M urea or DTT show a complete abolition of yield point, while those washed clean of urea and DTT prior to stretching approach native controls. Threads stretched in the presence of EDTA show no effect; however, preincubation of distal threads in EDTA for 24 h results in a loss of yield point if stretched in metal-deficient seawater and normal behavior in natural seawater. The results indicate that while protein unfolding and disruption of disulfide linkages or chelate complexes compromises the yield strength of distal byssal fibers, there is typically a rapid recovery in natural sea water.

Animals↗

PubMed2001

Reactivity of peptidyl-tyrosine to hydroxylation and cross-linking.

Tyrosine residues of neuroendocrine peptides are frequently the targets of oxidation reactions, one of which involves hydroxylation to peptidyl-3, 4-dihydroxy-phenyl-L-alanine (DOPA). The reactivity in vitro of peptidyl-DOPA in two neuroendocrine peptides, a neurotensin fragment (pELYENK) and proctolin (RYLPT), was investigated using ultraviolet-visible scanning spectrophotometry and matrix-assisted laser desorption ionization mass spectrometry following oxidation by tyrosinase and periodate. The peptides form covalently coupled dimers and trimers, and their masses are consistent with the presence of diDOPA cross-links. Lysine does not appear to participate in multimer formation because it is efficiently recovered in fragmentation ladders using subtilisin. While multimer formation in the neurotensin-derived peptide can be blocked effectively by adding N-acetyl-DOPA-ethylester to the reaction medium, the DOPA ethylester couples itself four to five times to each peptide.

Agaricales↗

PubMed2000

Cross-linking in adhesive quinoproteins: studies with model decapeptides.

Mytilus edulis foot protein-1 (mefp1) is a major component of the byssus, an adhesive holdfast in mussels. The recent report of 5, 5'-di(dihydroxyphenyl-L-alanine) (diDOPA) cross-links in byssus [McDowell et al. (1999) J. Biol. Chem. 274, 20293] has raised questions about the relationship of these to mefp1. About 80% of the primary structure of mefp1 consists of a tandemly repeated consensus sequence Ala(1)-Lys(2)-Pro(3)-Ser(4)-Tyr(5)-Pro(6)-Pro(7)-Thr(8)-Tyr(9)-Lys(10 ) with varying degrees of posttranslational hydroxylation to hydroxyprolines in positions 3, 6, and 7 and to DOPA in positions 5 and 9. Six natural or synthetic variants of this decapeptide were subjected to oxidation by tyrosinase or periodate. DOPA is the only residue to suffer losses in all oxidized peptides. Moreover, using MALDI TOF mass spectrometry, oxidized decapeptides all showed evidence of multimer formation and a mass loss of 6 Da per coupled pair of peptides. Multimer formation was inhibited by addition of DOPA-like o-diphenols, but addition of simple amines such as free Lys had no effect. The results are consistent with aryloxy coupling to diDOPA followed by reoxidation to diDOPA quinone. There are subtle but noteworthy variations, however, in multimer formation among the peptide congeners. Decapeptides with Pro(3) modified to trans-4-hydroxyproline do not form multimers beyond dimers; they also exhibit significant Lys losses following oxidation of DOPA. Moreover, in Ala-Lys-Hyp-Ser-Tyr-DiHyp-Hyp-Thr-DOPA-Lys, Tyr appears to be protected from oxidation by tyrosinase.

Adhesiveness↗

PubMed2000

The adhesive protein of Choromytilus chorus (Molina, 1782) and Aulacomya ater (Molina, 1782): a proline-rich and a glycine-rich polyphenolic protein.

The adhesive polyphenolic proteins from Aulacomya ater and Choromytilus chorus with apparent molecular masses of 135000 and 105000, respectively, were digested with trypsin and the peptides produced resolved by reversed phase liquid chromatography. About 5 and 12 major peptides were obtained from the protein of A. ater and C. chorus, respectively. The major peptides were purified by reverse-phase chromatography and the amino acid sequence indicates that both polyphenolic proteins consisted of repeated sequence motifs in their primary structure. The major peptides of A. ater contain seven amino acids corresponding to the consensus sequence AGYGGXK, whereas the tyrosine was always found as 3, 4-dihydroxyphenylalanine (Dopa), the X residue in position 6 was either valine, leucine or isoleucine, and the carboxy terminal was either lysine or hydroxylysine. On the other hand, the major peptides of C. chorus ranged in size from 6 to 21 amino acids and the majority correspond to the consensus sequence AKPSKYPTGYKPPVK. Both proteins differ markedly in the sequence of their tryptic peptides, but they share the common characteristics of other adhesive proteins in having a tandem sequence repeat in their primary structure.

Amino Acid Sequence↗

PubMed2000

Immunolocalization of Dpfp1, a byssal protein of the zebra mussel Dreissena polymorpha.

The zebra mussel is one of only a few freshwater bivalves known to produce a byssus. This fibrous, proteinaceous and highly cross-linked structure allows the mussel to attach to a variety of substrata and contributes to its notoriety as a major freshwater biofouling species. We have successfully expressed a full-length version of Dreissena polymorpha foot protein 1 (Dpfp1), a putative byssal thread precursor, and have used the recombinant protein as an antigen for polyclonal antibody production. Antisera obtained from rabbits immunized with recombinant Dpfp1 recognize the protein in western blots of extracts from foot tissue and byssal threads. On the basis of this evidence, we conclude that Dpfp1 is a byssal precursor protein manufactured and stored in the foot of the mussel. Immunohistochemical localization of Dpfp1 suggests that the protein is localized in secretory granules in a large gland surrounding the ventral groove of the foot. Only a subset of these glandular cells stockpiles the protein, implying that the zebra mussel foot is a complex organ capable of several distinct secretory activities involved in byssal thread formation. The uniform distribution of Dpfp1-containing cells suggests that the protein is a significant load-bearing component of zebra mussel byssal threads, although a more rigorous test of this hypothesis awaits ultrastructural localization of the protein in mature byssal threads.

Amino Acid Sequence↗

PubMed2000

In search of molecular dovetails in mussel byssus: from the threads to the stem.

We recently described the cDNA sequence for a unique collagenous protein, preCol-P, in the byssal threads of the marine mussel Mytilus edulis. The translated amino acid sequence encodes an unprecedented block-copolymer-like sequence with a central collagenous domain flanked by elastin-like sequences. Here, we report on the presence of two additional variants of preCol-P. The distribution of these variants in M. edulis foot tissue was examined by reverse transcription followed by polymerase chain reaction (RT-PCR) and in situ hybridization techniques. One of the variants, P33, exhibits a graded distribution with decreasing concentrations along the longitudinal axis of the foot. The second variant, P22, is expressed only at the base of the mussel's foot. In situ hybridization confirms the exclusive expression of preCol-P variant P22 in the stem gland. We propose that this variant may represent a molecular 'dovetail' between the proximal thread and the byssal stem, imparting extensibility and elastic recoil to the ring portion of the stem.

Animals↗

PubMed1999

Rotational echo double resonance detection of cross-links formed in mussel byssus under high-flow stress.

13C2H rotational echo double resonance NMR has been used to provide the first evidence for the formation of quinone-derived cross-links in mussel byssal plaques. Labeling of byssus was achieved by allowing mussels to filter feed from seawater containing L-[phenol-4-13C]tyrosine and L-[ring-d4]tyrosine for 2 days. Plaques and threads were harvested from two groups of mussels over a period of 28 days. One group was maintained in stationary water while the other was exposed to turbulent flow at 20 cm/s. The flow-stressed byssal plaques exhibited significantly enhanced levels of 5, 5'-di-dihydroxyphenylalanine cross-links. The average concentration of di-dihydroxyphenylalanine cross-links in byssal plaques is 1 per 1800 total protein amino acid residues.

Animals↗

PubMed1999

Reverse engineering of bioadhesion in marine mussels.

Marine mussels (Mytilus) are experts at bonding to a variety of solid surfaces in a wet, saline and turbulent environment. Bonding is rapid, permanent, versatile and protein-based. In mussels, adhesive bonding takes the form of a byssus--a bundle of extracorporeal threads--each connected to living tissues of the animal at one end and secured by an adhesive plaque at the other. We have investigated the composition and formation of byssal plaques and threads with the hope of discovering technologically relevant innovations in chemistry and materials science. All proteins isolated from the byssus to date share the quality of containing the unusual amino acid, 3,4-dihydroxyphenylalanine. This residue appears to have a dual functionality with significant consequences for adsorption and cohesion. On the one hand, it forms a diverse array of weaker molecular interactions such as metal chelates, H-bonds, and pi-cations: these appear to dominate in surface behavior (adsorption). On the other hand, 3,4-dihydroxyphenylalanine and its redox couple, dopaquinone, can mediate formation of covalent cross-links among byssal proteins (cohesion). One of the challenges in making functional biomimetic versions of byssal adhesion is to understand how these two reactivities are balanced.

Adhesives↗

PubMed1998

Structure and mucoadhesion of mussel glue protein in dilute solution.

Purified mussel adhesive protein mefp-1 (Mytilus edulis foot protein 1) has been studied regarding its state of oligomerization and gross conformation in dilute solution. Sedimentation equilibrium in the analytical ultracentrifuge of a dilute solution of protein (0.4 mg/mL) in acetate buffer at pH 4.5 and I = 0.10 M yielded an apparent molecular weight (whole distribution weight average, Mw, app) of 114 000 +/- 5000 via the "M" procedure, a value in almost exact agreement with the monomeric molecular weight obtained by MALDI mass spectrometry. At this low concentration, it is reasonable to assume thermodynamic ideality, i.e., Mw,app approximately Mw. This result, together with plots of point weight average apparent molecular weight versus concentration for three different loading concentrations (0.4, 0.8, 1.0 mg/mL), clearly demonstrates that this protein is essentially monomeric in dilute solution. Sedimentation velocity experiments yielded an estimate of the sedimentation coefficient s020,w = 2.34 +/- 0.17 S, which for M = 110 000 gives a frictional ratio f/f0 = 3.2 +/- 0.3. The interpretation of this, in terms of an extended rather than globular conformation for the structure of mefp-1 in dilute solution, is considered, within plausible limits of molecular hydration, and models for the structure in solution are considered, in light of the thermodynamic nonideality behavior of these molecules and previously published circular dichroism data. The significance of these observations in terms of the bioadhesive properties of mefp-1 is described, and the very strong interaction in dilute solution with a mucin glycoprotein is demonstrated.

Adhesiveness↗