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J H Waite

Publications and source records attributed to J H Waite.

At least 55 records · Page 3Linked to original sources
PubMed1993

The byssus of the zebra mussel, Dreissena polymorpha. I: Morphology and in situ protein processing during maturation.

The zebra mussel, Dreissena polymorpha, owes its notoriety as a biofouler to its adhesive skills and opportunism. Adhesion by the adult mussel to hard substrata is mediated by a nonliving extracorporeal structure called the byssus, which is superficially similar to the byssus of marine mussels in that it consists of a tight bundle of sclerotized threads tipped by adhesive plaques. Juvenile zebra mussels secrete a homologous structure on settlement, but they also employ an elongated belaying byssus while climbing that consists of an elastic, mucous filament anchored at irregular intervals by a byssal thread and plaque. This multiply anchored belaying line can be 20 to 30 times the mussel length. Histochemical tests show that the thread and plaque of both kinds of byssus contains a complex distribution of proteins that are subject to chemical processing after secretion. This processing may result from the formation of crosslinks following the catecholoxidase-catalyzed oxidation of peptidyl 3,4-dihydroxyphenylalanine during sclerotization.

Aging↗
PubMed1993

The byssus of the zebra mussel, Dreissena polymorpha. II: Structure and polymorphism of byssal polyphenolic protein families.

The byssus of the zebra mussel Dreissena polymorpha is the key element of its adhesive strategy. It consists of a bundle of threads tipped by adhesive plaques and attached to the mussel at the base of its byssal-synthesizing organ, the foot. Two polyphenolic protein precursors of the byssus have been purified from the foot. These precursors, Dpfp-1 and Dpfp-2 (Dreissena polymorpha foot protein), with apparent M(r) values of 76 and 26 K, respectively, contain 3,4-dihydroxyphenylalanine (DOPA) integrated into their primary sequence, but differ from previously characterized polyphenolic proteins from marine mussels. The related quagga mussel (Dreissena spp.?) has homologous proteins with significantly different compositions. The zebra mussel DOPA proteins are tandemly repetitive with unique oligopeptide motif sequences, contain tryptophan, and are O-glycosylated primarily on threonine residues. Galactosamine was the only carbohydrate detected after hydrolysis. Dpfp-1 constitutes a polymorphic family of polypeptides with, unusually, an acidic range of pI values between 5.3 and 6.5. The detection of carbohydrate in the thread and in the juncture between thread and plaque suggests that these two proteins are localized in those regions where they may function as lacquers or structural elements.

Amino Acid Sequence↗
PubMed1992

Cement precursor proteins of the reef-building polychaete Phragmatopoma californica (Fewkes).

Two distinctive 3,4-dihydroxyphenyl-L-alanine-(DOPA-) containing proteins (Pc-1 and Pc-2) have been isolated and partially characterized from the thorax of the reef-building sabellariid Phragmatopoma californica. They are the first such reported from the phylum Annelida. The proteins are presumed to be soluble precursors of the quinone-tanned cement used to bind particulate materials in the construction of the tubes that serve as habitats for the worms. The proteins have apparent molecular weights ranging from 18,000 to 20,000 and isoelectric point greater than or equal to 8.0. Both proteins consist of repeated sequence motifs in their primary structure. Pc-1 has repeats of (XGGY*GY*GAK) where X = V, L, I, AA, or KV, and Y* is DOPA or tyrosine. Pc-2, in contrast, appears to have repeats of (X1-[GGY*]n-[GA]m-X2-[HP(A)V]p-HK) where X1 can be AL, A, or F; X2 can be WG or absent; n and m can be 1 or 2, and p = 0-2. Both protein families appear to share the same C-terminal sequence ALGGY*GAGA. Of the DOPA-containing proteins characterized from other phyla, Phragmatopoma cement precursors most resemble those from the liver fluke Fasciola hepatica and the mussel Trichomya hirsuta.

Amino Acid Sequence↗
PubMed1992

Discovery of bone gamma-carboxyglutamic acid protein in mineralized scales. The abundance and structure of Lepomis macrochirus bone gamma-carboxyglutamic acid protein.

The mineralized scale of the freshwater sunfish Lepomis macrochirus (bluegill) contains a Gla protein. The protein was identified in extracts of scale by a new colorimetric assay for Gla-containing proteins. The protein was purified by gel filtration chromatography followed by reversed phase high performance liquid chromatography (HPLC). Several tests establish the identity of scale Gla protein and bone Gla protein (BGP). First, the proteins exhibit identical mobilities on electrophoresis and by reversed phase HPLC. Second, they have identical amino-terminal amino acid sequences. Finally, identical peptides are generated by proteolytic digestion. The 45-residue amino acid sequence of the bone Gla protein from L. macrochirus has a high sequence homology with swordfish, as well as homology to mammalian bone Gla protein. The BGP of bluegill shares with swordfish BGP a truncated NH2 terminus and an extended COOH terminus. These features may be unique to fish, as they have not been observed in terrestrial vertebrates. The bluegill BGP is the first vitamin K-dependent protein to contain a non-gamma-carboxylated residue to the NH2-terminal side of all of its Gla residues. In all other vitamin K-dependent proteins, Gla always appears to the NH2-terminal side of the first Glu. The implications of this result are discussed. The bluegill rib bone is curiously enriched in BGP, as are other mineralized tissues of this species. One hypothesis is that this may be due to the acellular nature of the bone in this species. The abundance of BGP in the bones of this fish may provide clues to the unknown function of this bone protein.

Amino Acid Sequence↗
PubMed1992

Eggshell precursor proteins of Fasciola hepatica, I. Structure and expression of vitelline protein B.

Antibody raised against the major eggshell protein of Fasciola hepatica (vitelline protein B, vpB) is employed to isolate cDNAs from an expression library and to localize the protein in whole worms. Two cDNAs corresponding to the protein are homologous through the N-terminal and C-terminal coding regions and widely divergent internally. No repeated regions are apparent and no significant sequence homology is seen with chorion proteins of other genera although the amino acid composition closely reflects that of other chorion proteins. Microheterogeneity observed in the vpB is due to the presence of multiple coding sequences, transcripts and a gradient of post-translational modification. Relative transcription of the vpB mRNA throughout the female reproductive tract is demonstrated.

Amino Acid Sequence↗
PubMed1992

Eggshell precursor proteins of Fasciola hepatica, II. Microheterogeneity in vitelline protein B.

At least 3 structural protein precursors of the eggshell are synthesized and stockpiled in the extensive vitelline cells of the liver fluke Fasciola hepatica L. One of these, vitelline protein B, consists of a closely related family of proteins that owes its apparent electrophoretic heterogeneity to variations in the Tyr to DOPA conversion as well as to subtle variations in the primary sequence. The efficiency of the Tyr to DOPA conversion ranges from a maximum of about 90% to a minimum of 55% in the protein. Trypsin digestion in borate buffer at pH 8 was used to produce DOPA-peptides for sequencing. Notably, trypsin does not cleave Arg/Lys-DOPA sequences at borate concentrations greater than 0.15 M. Peptides with DOPA-containing sequences most frequently have flanking amino acids such as Lys, Ser, or Asp on the N-terminal side and Gly or Asp on the C-terminal side. All protein variants fall within a narrow molecular weight range (30-33 kDa), a pI range of 6.9 to 8.3, and the collective majority would appear to share a common N-terminal sequence up to residue 28. The results suggest some combination of the following: variations in post-translational hydroxylation, alternative post-transcriptional splicing and/or the existence of multiple gene copies of eggshell precursors. The latter have been shown to occur in the blood fluke Schistosoma mansoni [15].

Amino Acid Sequence↗
PubMed1991

alpha,beta-Dehydro-3,4-dihydroxyphenylalanine derivatives: potential schlerotization intermediates in natural composite materials.

Proteins containing the post-translationally modified amino acid L-3,4-dihydroxyphenylalanine (DOPA) undergo autosclerotization as a means of assuring cohesive resilience in many structural matrices found in nature. To explore the chemical mechanism of sclerotization, we examined the oxidation products of relatively simple analogs of a peptidyl DOPA residue, such as N-acetylDOPA ethyl ester and N-acetyldopamide, together with those of several oligopeptides. Oxidation, induced by either of two catecholoxidases or by sodium periodate, resulted in the Lewis base catalyzed formation of derivatives of the unusual amino acid 3,4-dihydroxy-alpha,beta-dehydroDOPA (delta DOPA). The N-acetyl delta DOPA ethyl ester representative of this group of derivatives was characterized by NMR and uv spectroscopy. A variety of peptides developed analogous uv spectra upon oxidation. A similar reaction was observed upon oxidation of 3,4-dihydroxyphenylpropanoic (dihydrocaffeic) acid, but not after oxidation of N-acetyldopamine. Evidence is presented that this conversion is the result of a rearrangement of the DOPA quinone moiety to its delta DOPA tautomer, and that this tautomerization can be a dominant fate for peptidyl DOPA quinone, provided a Lewis base catalyst is available and competing reactions are minimized. Formation of delta DOPA in natural or synthetic polymers would increase the variety of crosslinks available to sclerotizing matrices. delta DOPA has been found in naturally occurring oligopeptides isolated by other workers from several marine species.

Amino Acid Sequence↗
PubMed1991

alpha,beta-Dehydro-3,4-dihydroxyphenylalanine derivatives: rate and mechanism of formation.

The amino acid L-3,4-dihydroxyphenylalanine (DOPA), when present in the primary sequence of proteins, does not form melanin upon oxidation to the quinone, since its amine moiety participates in a peptide bond and cannot undergo internal cyclization. Instead, peptidyl DOPA quinone is available for other reactions. We have investigated the oxidation chemistry of a low molecular weight peptidyl DOPA analog, N-acetylDOPA ethyl ester (NAcDEE), and have shown that a major product of oxidation is an unsaturated DOPA derivative, N-acetyl-alpha,beta-dehydroDOPA ethyl ester (NAc delta DEE) (see companion paper, Rzepecki et al., Arch. Biochem. Biophys. (1991) 285, 17-26). In the present study, we have explored kinetic and mechanistic features of the conversion of NAcDEE to NAc delta DEE and found that the reaction requires: (i) oxidation of NAcDEE to the quinone, (ii) the presence of a Lewis base as a catalyst (phosphate anion was the best of those tried in the pH range 6.0-8.0), and (iii) prevention of competing reactions such as Michael additions. Conversion efficiencies in the presence of Lewis bases ranged between 12 and 19% at pH 8.0 and 35 and 90% at pH 6.0. At least two separate reaction mechanisms appeared necessary to explain the kinetic data: (i) a pseudo-first-order mechanism at pH 6.0 and above, and (ii) an additional second-order mechanism at higher pH which involved both NAcDEE catechol and quinone. The apparent pseudo-first-order rate constants increased with pH from 2.36 X 10(-4) s-1 at pH 6.0 to about 30 X 10(-4) s-1 at pH 8.0 in 0.1 M sodium phosphate. Tautomerization of DOPA quinone to dehydroDOPA may thus be a factor in the sclerotization of natural structures incorporating DOPA containing proteins.

Buffers↗
PubMed1991

Detection of peptidyl-3,4-dihydroxyphenylalanine by amino acid analysis and microsequencing techniques.

3,4-Dihydroxyphenylalanine (DOPA) is an amino acid that occurs naturally in the primary sequence of many proteins and peptides. Detection of peptidyl-DOPA, however, can be elusive. This is due (i) to its coelution with leucine on most of the ion exchangers used in amino acid analysis and (ii) to the coelution of phenylthiohydantoin (PTH)-DOPA with PTH-alanine during routine C18 reversed-phase HPLC following automated Edman degradation. By application of appropriately timed temperature and/or gradient modifications during chromatography, DOPA and its PTH derivative can be adequately resolved for detection by both amino acid analysis and gas-phase sequencing.

Amino Acid Sequence↗
PubMed1991

Molecular diversity of marine glues: polyphenolic proteins from five mussel species.

Adhesive polyphenolic proteins have been purified and characterized from the feet of five marine mussels (Brachidontes exustus, Modiolus modiolus squamosus, Mytella guyanensis, Septifer bifurcatus, and Trichomya hirsuta). All five proteins contain high levels of 3,4-dihydroxyphenylalanine (DOPA), lysine, glycine, and serine or threonine. All but B. exustus also contain high levels (> or = 10%) of proline or 4-hydroxyproline. The polyphenolic proteins of all the mussels have repeated sequences of the motif X1-Y*-X2-Y*-X3-K, where Y* denotes tyrosine or DOPA. In two species (S. bifurcatus and B. exustus), X2 represents 3 amino acids (frequently glycine) and X3 is absent. M. guyanensis is similar except that X2 is reduced to 2 amino acids. In T. hirsuta and M. m. squamosus, however, X2 is absent and X3 occurs as alanine or hydroxyproline. All proteins share approximately equimolar proportions of tyrosyl- and lysyl-derived residues. Although all of the mussels examined thus far are adhesively opportunistic with respect to substratum type, a rigidly invariant sequence does not appear to be necessary for achieving this.

Amino Acid Sequence↗
PubMed1990

Marine adhesive proteins: natural composite thermosets.

Marine environments are severely challenging for the performance and durability of synthetic adhesives. Factors commonly associated with adhesive failure are weak boundary layers (water, oxides), adhesive erosion and swelling. For many permanently attached marine organisms such as barnacles, mussels, oysters, etc., however, underwater adhesion is 'business-as-usual'. Knowledge about the chemistry and bioprocessing of these marine adhesives will provide profound insights for the evolution of a new generation of environmentally safe, water-resistant adhesives. Despite their apparent structural diversity, marine adhesives are essentially analogous to composite thermosets, that is, the adhesive consists of fibre, filler and catalyst molecules that are dispersed in a cross-linked resin rendering it resistant to heat and solvents. The fibres and fillers in these composites are variable. e.g. collagen, fibroin, chitin present as fibres, and sand, shell, air and water present as fillers. The precured resins of seven organisms including members of the Mollusca, Annelida, and Platyhelminthes have now been isolated and partially sequenced. These are proteins with basic isoelectric points, high levels of the amino acid 3,4-dihydroxyphenyl-L-alanine (DOPA), and an extended, flexible conformation. The DOPA functional group in particular is thought to play a key role in (a) the chemisorption of these polymers to surface underwater, and (b) covalent cross-linking or setting of the adhesive, the latter reaction catalysed by the enzyme catecholoxidase. Much more needs to be done to explore the details of the adhesive processing and delivery strategies used by these organisms.

Adhesiveness↗
PubMed1990

The phylogeny and chemical diversity of quinone-tanned glues and varnishes.

1. 3,4-Dihydroxyphenyl-L-alanine (DOPA)-containing proteins are widely distributed throughout the animal kingdom and appear to serve chiefly as waterproof adhesives and varnishes. 2. The unique chemical and physical stability of these adhesives and varnishes is imparted by quinone-tanning, an oxidative process that leads to the polymerization of DOPA-containing and other proteins. 3. Recent advances in the biochemistry of DOPA-containing proteins suggest that most consist of tandemly repeated sequence motifs. Each motif contains DOPA, a basic amino acid (usually lysine), and abundant glycine or proline. 4. The DOPA residues undergo catechol oxidase-catalyzed conversion to o-quinones at the onset of quinone-tanning. 5. The complexity of quinone chemistry is discussed with regard to quinone-tanning.

Adhesives↗
PubMed1989

A histidine-rich protein from the vitellaria of the liver fluke Fasciola hepatica.

The vitellaria are an extensive network of glandular cells and ducts distributed throughout the peripheral tissues of the liver fluke Fasciola hepatica. Eggshell precursor proteins are produced and stockpiled in the vitelline cells of mature flukes. Vitelline protein C has an extraordinary composition: the amino acid 3,4-dihydroxyphenyl-L-alanine (DOPA) and histidine each comprise about 20% of the residues, while glycine represents 41-42% in all variants of what appears to be a microheterogeneous protein family. Protein C has an apparent molecular weight of 16,000-17,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Although the protein appears homogeneous following polyacrylamide gel electrophoresis in Tris-glycine with SDS and a acetic acid-urea, electrophoresis in borate, however, suggests that the vitelline protein consists of four or more closely related proteins weighing from 16,000 to 18,500. Isoelectric focusing of the protein family in the presence of 8 M urea resolved only two species having pI values of 6.89 and 6.99. A single N-terminus having the sequence H-H-W-D-G-DOPA-G-DOPA-G was detected. The primary structure of vitelline protein C is characterized by a repeated motif consisting of (G-X)n, where X is Ser, DOPA, or His. Most of the His occurs as G-H repeats in a pepsin-resistant fragment of the protein. Previously, a 31-kDa protein, representing up to 6% of the total protein in the fluke, was reported [Waite, J. H., & Rice-Ficht, A (1987) Biochemistry 26, 7819-7825] to contain significant levels of DOPA.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1989

The glue protein of ribbed mussels (Geukensia demissa): a natural adhesive with some features of collagen.

The Atlantic ribbed mussel Geukensia (Modiolus) demissa attaches itself to the roots of cord grass and other hard objects in tidal salt marshes by spinning adhesive byssal threads. The precursor of a protein apparently present in the adhesive plaques of the threads was isolated in quantity from the foot of the mussel. The protein has an apparent molecular weight of 130,000, a pI of 8.1, and contains a high proportion of Gly, Glu/Gln, Lys and 3,4-dihydroxyphenyl-L-alanine (DOPA). Sequence of tryptic peptides suggests a pattern of repeated motifs, such as: Gly--DOPA--Lys, and X--Gly--DOPA--Y--Z--Gly--DOPA/Tyr--Lys, where X is Thr or Ala in octapeptides and Gln--Thr in nonapeptides. Y is variable, but more often than not hydrophobic; and Z is frequently Pro or 4-trans-hydroxyproline (Hyp). The presence of Pro--Gly and Hyp--Gly sequences of delta-hydroxylysine in the protein is reminiscent of typical collagens; however, the protein is not labile to clostridial collagenase, nor does collagen cross-react with antibodies raised against the mussel protein. Unlike typical collagens, Gly probably occurs only at every 4th or 5th residue in this unusual mussel protein.

Adhesives↗
PubMed1989

A chromogenic assay for catecholoxidases based on the addition of L-proline to quinones.

The coupling reaction between L-proline and the quinone products of the oxidation of various catechols serves as a sensitive assay for catecholoxidases. The chromogenic products, 4-N-prolyl-o-quinones, were unique and stable over the course of the reaction. The spectra of these adducts typically had two absorbance maxima, in the ranges 309-340 and 526-540 nm. Assay conditions in which the oxidation of catechols was rate limiting were developed, and initial rates of reaction, monitored spectrophotometrically at the lambda max of the adducts, showed improved initial linearity when compared with the direct spectrophotometric determination of quinone formation. The molar extinction coefficients (epsilon) of a number of adducts ranged between 5310 and 9630 M-1 cm-1, about five- to sevenfold greater than those of the corresponding quinones. Since 2 mol catechol must be oxidized to their quinones to generate 1 mol of adduct, this assay improves catecholoxidase detection sensitivity by approximately three- to fourfold compared with direct estimation of quinone formation.

Benzoquinones↗
PubMed1989

Mussel glue protein has an open conformation.

Both native glue protein from marine mussels and a synthetic nonhydroxylated analog were analyzed by far-uv CD under a variety of conditions. Analysis of the CD spectra using various models strongly suggest a primarily random coil structure for both forms of the protein, a fact also supported by the absence of spectral change for the glue protein upon dilution into 6 M guanidine hydrochloride. The nonhydroxylated analog, which consists of 20 repeats of the peptide sequence Ala-Lys-Pro-Ser-Tyr-Pro-Pro-Thr-Tyr-Lys, was further characterized by enzyme modification using mushroom tyrosinase. Enzymatic hydroxylation of tyrosines was found to be best fit by a model containing two rate constants, 5.6 (+/- 0.6) X 10(-3) and 7.2 (+/- 0.3) X 10(-2) min-1. At equilibrium, HPLC analysis of digests showed nearly 100% conversion of Tyr-9 and only 15 to 35% conversion of Tyr-5. The Chou and Fasman rules for predicting structure were applied to the repeat sequence listed above. The rules predict the absence of alpha helix and beta pleated sheets in the structure of this peptide. On the other hand, beta turns are predicted to be present with Tyr-5 being in the region of highest probability. These data suggest that the protein in solution has only a small amount of secondary structure.

Amino Acid Sequence↗
PubMed1987

Presclerotized eggshell protein from the liver fluke Fasciola hepatica.

Trematode parasites protect their eggs with a tough tanned eggshell. Eggshell precursor proteins are synthesized and stockpiled within the extensive vitellaria of the animal. A major eggshell precursor protein with an apparent molecular weight of 31,000 and pI of 7.4 was isolated from the vitellaria of Fasciola hepatica. This protein, which represents 6-7% of the total protein in mature Fasciola, is unique in containing rather high levels of the amino acid 3,4-dihydroxyphenylalanine (DOPA), i.e., 110 residues per 1000. Other prominent amino acids are glycine, aspartic acid, and lysine. A prominent DOPA-containing tryptic peptide derived from eggshell precursor protein has the sequence Gly-Gly-Gly-DOPA-Gly-Gly-DOPA-Gly-Lys. DOPA residues disappear during the maturation of the eggshell and by treatment in vitro with mushroom polyphenol oxidase. This disappearance may be related to the formation of cross-links in the eggshell protein.

Amino Acid Sequence↗