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D A Parry

Publications and source records attributed to D A Parry.

At least 37 records · Page 2Linked to original sources
PubMed1998

Hard alpha-keratin intermediate filament chains: substructure of the N- and C-terminal domains and the predicted structure and function of the C-terminal domains of type I and type II chains.

The quantity of sequence data now available for both Type I and Type II hard alpha-keratin IF proteins makes it possible to analyze their N- and C-terminal domains and ascertain features of likely structural and/or functional importance. The N-terminal domains of both chain types can be divided into acidic (NA) and basic (NB) subdomains, where NA is 29 and 34 residues long, respectively, for Type I and II chains and is located immediately adjacent to the end of the rod domain. NB constitutes the remainder of the N-terminal domain and is about 27 and 70 residues long for the two chain types, respectively. The glycine residue contents, however, are high in NA(I) and NB(II), but low in NA(II) and NB(I). Subdomain NB(II) contains four consecutive nonapeptide quasirepeats of the form GGGFGYRSX. The C-terminal domain of Type I chains, termed C(I), is characterized by a PCX motif repeated 10 times, 7 of them contiguously. From an analysis of the conformation of like peptides from crystal structures it has been shown that this region will probably adopt a polyproline II left-handed helical structure with three residues per turn. In contrast, the C-terminal domain of Type II hard alpha-keratin chains (known as C(II)) contains a periodic distribution of hydrophobicities that, together with other predictive techniques, allow its conformation (a twisted four-stranded antiparallel beta-sheet) to be predicted with some degree of confidence. In addition, it is possible to suggest two partners with which this domain will interact. The first is with segment L12 in the rod domain and the second is with another C(II) domain in an antiparallel neighboring molecule. The latter possibility appears most likely. In either case the aggregation would likely serve to stabilize the molecular assembly through the interaction of two beta-sheets via their apolar faces and, in so doing, would position a number of cysteine residues in external positions that would allow them to form a number of covalent disulfide bonds with other molecules.

Amino Acid Sequence↗
PubMed1998

A conserved C-terminal assembly region in paramyosin and myosin rods.

The assembly of myosin and paramyosin into filaments in muscle has been shown to depend in part on the interactions of regular periodic patches of charge on the surface of the rod regions of these alpha-helical coiled-coil proteins. It has also been known for some time that a relatively small region near the C-terminus of both molecules is critical for both solubility and assembly. This domain appears to function as a modulator of assembly in both proteins. Recently, a specific 29-residue region in the C-terminus of human fast muscle myosin rod has been shown to be essential for filament formation, and this sequence has been shown to be present in other vertebrate and invertebrate myosins. We show here that paramyosin also displays this specific conserved domain. Moreover, we have found that this domain is part of a longer distinctive region in both paramyosin and myosin: this region lacks the periodic variation in charge found in the rest of both coiled coils, has a unique charge profile, a relatively neutral total charge, and a high proportion of large apolar residues in surface positions. These results may be useful in designing site-directed mutagenesis studies to identify the target regions on neighboring molecules which interact with this C-terminal domain and so establish the mechanism of its function.

Actin Cytoskeleton↗
PubMed1998

Identification of multiple forms of 180-kDa ribosome receptor in human cells.

Herein, we describe the analysis and mapping of cDNA clones encoding variant forms of the human homolog of the canine 180-kDa ribosome receptor (p180). One form, similar to the chicken ES/130 homolog, possesses a large uninterrupted C-terminal region composed predominantly of heptad repeats predicted to form an alpha-helical double-stranded coiled-coil rod. Other forms contain in addition a 10-amino acid consensus motif, NQGKKAEGAQ, repeated up to 54 times in tandem close to the N-terminus. Such repeats in canine p180 represent a ribosome-binding domain. The cDNA hybridized to a major 6-kb transcript in all tissues examined, where very high expression was observed in tissues that carry out a high level of secretion such as pancreas, liver, and placenta. The ES130/p180 gene was mapped to chromosome 20p12, and a potential pseudogene appears to reside on chromosome 7. In summary, the data suggest that p180 exists in humans in different forms because of complete removal of tandem repeats, or partial intraexonic splicing, creating different repeat lengths with potentially novel ribosome-binding characteristics.

Alternative Splicing↗
PubMed1998

Exercise-related alterations in crimp morphology in the central regions of superficial digital flexor tendons from young thoroughbreds: a controlled study.

Injury to the core of the mid-metacarpal region of the superficial digital flexor tendon in Thoroughbred racehorses is a very frequent but poorly understood condition. It has been suggested that subclinical changes induced by galloping exercise weaken the collagen in this region of the tendon, predisposing it to rupture. The longitudinally arranged collagen fibrils in tendon follow a planar waveform, termed the crimp. Fibril bundles with a smaller crimp angle fail at a lower level of strain than those with a larger crimp angle. This study tested the hypothesis that a specific 18 month exercise programme would result in significant reduction of collagen fibril crimp angle and period length in the core region of the superficial digital flexor tendon of young Thoroughbreds (21 +/- 1 months), compared to the normal change in these parameters with age. Central region crimp angle and length were significantly lower in exercised horses than in control horses (P < 0.05). The crimp angle was significantly lower in this central region than in the peripheral region of the tendon in 4 of the 5 exercised horses, as was the crimp length in 3 of the 4 horses. The crimp angle in the peripheral region was significantly greater in exercised horses than in the controls (P < 0.05), which may indicate functional adaptation due to differing mechanical environment between the 2 tendon regions. The results of this study supported previous evidence that galloping exercise modifies normal age-related changes in crimp morphology in the core of the superficial digital flexor tendon. Such changes are indicative of microtrauma and would be detrimental to tendon strength.

Aging↗
PubMed1998

Effects of training on collagen fibril populations in the suspensory ligament and deep digital flexor tendon of young thoroughbreds.

OBJECTIVE: To determine the effect of a specific galloping exercise regimen on collagen fibril mass-average diameters (MAD) in the deep digital flexor tendon (DDFT) and suspensory ligament (SL) of young Thoroughbreds. ANIMALS: 12 Thoroughbred fillies, 21 +/- 1 (mean +/- SD) months old. PROCEDURE: 6 horses underwent a specific 18-month treadmill training program involving galloping exercise. The remaining 6 horses served as controls, undertaking low-volume walking exercise over the same period. Sections were excised from the midpoint of the DDFT and SL, and small strips were dissected from central and peripheral locations for each structure. Fibril diameters were measured from micrographs of transverse ultrathin sections, using a computerized image analysis program. An MAD value was calculated for the central and peripheral regions of the DDFT and SL for each horse. Values for both regions were compared between exercised and control horses. RESULTS: The MAD did not change significantly with exercise for either the DDFT or the SL. CONCLUSION: Loading of the DDFT as a result of this exercise regimen was not sufficient to stimulate collagen fibril hypertrophy, in keeping with current data that indicate this tendon, compared with the SL and superficial digital flexor tendon (SDFT), is subjected to low loads. Microtrauma, in terms of reduction in fibril MAD, may have occurred in the SL at a site different from that sampled. Another possibility is that, between the trot and the gallop, loading of the SL does not increase to the same extent as that of the SDFT.

Animals↗
PubMed1997

Periplakin, a novel component of cornified envelopes and desmosomes that belongs to the plakin family and forms complexes with envoplakin.

The cornified envelope is a layer of transglutaminase cross-linked protein that is assembled under the plasma membrane of keratinocytes in the outermost layers of the epidermis. We have determined the cDNA sequence of one of the proteins that becomes incorporated into the cornified envelope of cultured epidermal keratinocytes, a protein with an apparent molecular mass of 195 kD that is encoded by a mRNA with an estimated size of 6.3 kb. The protein is expressed in keratinizing and nonkeratinizing stratified squamous epithelia and in a number of other epithelia. Expression of the protein is upregulated during the terminal differentiation of epidermal keratinocytes in vivo and in culture. Immunogold electron microscopy was used to demonstrate an association of the 195-kD protein with the desmosomal plaque and with keratin filaments in the differentiated layers of the epidermis. Sequence analysis showed that the 195-kD protein is a member of the plakin family of proteins, to which envoplakin, desmoplakin, bullous pemphigoid antigen 1, and plectin belong. Envoplakin and the 195-kD protein coimmunoprecipitate. Analysis of their rod domain sequences suggests that the formation of both homodimers and heterodimers would be energetically favorable. Confocal immunofluorescent microscopy of cultured epidermal keratinocytes revealed that envoplakin and the 195-kD protein form a network radiating from desmosomes, and we speculate that the two proteins may provide a scaffolding onto which the cornified envelope is assembled. We propose to name the 195-kD protein periplakin.

Amino Acid Sequence↗
PubMed1997

Protein chains in hair and epidermal keratin IF: structural features and spatial arrangements.

Over the past decade the progress made in characterising the structural hierarchy of both the hard and the epidermal keratin intermediate filaments has exceeded all expectations. The origin of much of this progress can be traced back to the quantity of amino acid sequence data that became available in the early/mid 1980s, and their interpretation in terms of a heterodimeric molecular structure. Subdomains were subsequently identified in both the rod and terminal domains, and now the roles of most of these have been determined in principle, if not yet fully in detail. TEM and STEM, together with very revealing crosslinking analyses have also allowed details to be determined of the mechanism by which molecules assemble into oligomers and oligomers into IF. It remains for the three-dimensional packing of keratin molecules in the IF to be elucidated, but even here progress is being made. A particularly exciting development over the last two or three years has been the establishment of the link between keratinopathies and single point nucleotide mutations in keratin genes. Furthermore, the clustering of mutation sites in regions involved in a key structural mode of molecular aggregation has provided, for the first time, an understanding of keratin diseases at the molecular level.

Amino Acid Sequence↗
PubMed1997

Intermediate filament structure: hard alpha-keratin.

Structurally there are four classes of intermediate filaments (IF) with distinct but closely related axial organisations. One of these, hard alpha-keratin IF, has been studied to clarify several apparently exceptional features which include the number of molecules in the IF cross-section and the mode by which the axial organisation of its constituent molecules is stabilised. Using the dark-field mode of the STEM at the Brookhaven National Laboratory (USA) mass measurements were obtained from unstained IF isolated from hair keratin. The data thus obtained show that the number of chains in cross-section is about 30 (+/-3: standard deviation) and is very similar to the numbers determined in previous STEM experiments for the dominant filament type in other classes of IF (about 32). Furthermore, re-analysis of the low-angle equatorial X-ray diffraction pattern reveals, in contrast to earlier work, solutions that are compatible with the number of chains in cross-section indicated by the STEM data. The absence of the head-to-tail overlap between parallel molecules characteristic of most of IF may be compensated in hard alpha-keratin by a network of intermolecular disulfide bonds. It is concluded that native IF of hard alpha-keratin and desmin/vimentin--and probably many other kinds of IF as well--contain about 32 chains in cross-section, and that the axial structures of these various kinds of IF differ in small but significant ways, while generally observing the same basic modes of aggregation.

Animals↗
PubMed1997

Exercise modifies the age-related change in crimp pattern in the core region of the equine superficial digital flexor tendon.

One of the current concepts with regard to equine superficial digital flexor tendonitis is that cumulative subclinical microscopic damage weakens the structure, predisposing the tendon to partial or complete rupture. This microtrauma is likely to affect the waveform or crimp of the collagen fibrils, which are the units of tensile strength. Collagen fibril crimp morphology characteristics were determined in superficial digital flexor tendons of 18 horses presented for necropsy. Horses were separated into exercised (n = 9) and non-exercised groups (n = 9), based on recent function. Five of the eight exercised horses below the age of 10 years showed a significantly lower crimp angle in the central region of the tendon in comparison with the periphery. Three of those five animals also showed a significantly lower crimp period length in the centre. No non-exercised horses in this age group showed such regional differences. Horses classified as non-exercised may have Horses classified as non-exercised may have undergone competitive galloping activity at an earlier age, implying that changes in central region crimp morphology did not occur at that time, or had reverted to normal values in the intervening period. A lower crimp angle in the core region in comparison with the periphery is abnormal in animals younger than 10 years, on the basis of previous data obtained from wild horses. It is hypothesised that imposed exercise regimens involving galloping modify the normal age-related reduction of crimp angle in the tendon core, probably as a result of the increased number of rapid high-strain cycles experienced by the collagen fibrils.

Journal Article↗
PubMed1997

Age-related differences in collagen crimp patterns in the superficial digital flexor tendon core region of untrained horses.

OBJECTIVE: To measure collagen fibril crimp angles and lengths as well as collagen fibril mass-average diameters in central and peripheral regions of the superficial digital flexor tendon of wild horses, to ascertain any age-related changes in either region in the absence of imposed galloping exercise. DESIGN: Measurements from a random cull of wild horses. SAMPLE POPULATION: Superficial digital flexor tendon samples were taken from 23 wild horses ranging in age from two to ten years. PROCEDURE: Horses were divided into 'young' (< 5 years, n = 10), 'middle-aged' (5 to < 10 years, n = 9) and 'old' groups (10+ years, n = 4) and the mean crimp angle, mean crimp length and collagen fibril mass-average diameter calculated for the central region, and for the peripheral region of each group. Differences between groups and regions were analysed using two-tailed t tests. RESULTS: The crimp angle for the central region was found to decrease with age, so that in old horses it was smaller than that for the tendon periphery (P, 0.05). The crimp angle for the latter region did not alter significantly with age. Crimp period lengths and collagen fibril mass-average diameters did not show significant changes with age in either region. CONCLUSIONS: Reduction of the crimp angle in the core of the superficial digital flexor tendon occurs normally with age, as tendons of older animals would have undergone a higher number of loading cycles. It is possible that athletic training increases the frequency and/or the magnitude of high loading cycles experienced by the tendon, and may accelerate and worsen the normal load-related ageing process in the superficial digital flexor tendons of young performance horses, particularly in the central regions where lesions usually occur.

Aging↗
PubMed1997

Comparison of collagen fibril populations in the superficial digital flexor tendons of exercised and nonexercised thoroughbreds.

This study was undertaken to test the hypothesis that collagen fibrils, the submicroscopic units of strength in tendon, would hypertrophy in response to a specific defined training programme. Fibril diameters were measured in central and peripheral regions of superficial digital flexor tendon (SDFT) samples from five 18-month-old horses which underwent a subsequent 18 month training programme and 6 age- and sex-matched controls. Central region fibrils from the trained horses had a mass-average diameter (MAD) of 105.3 nm, which was significantly lower (P < 0.01) than that of 131.7 nm for the same region in the control horses. This reduction in fibril diameter in the region of tendon which is predisposed to injury was interpreted as evidence of microtrauma, as it implies the region is weakened by the training regimen. Repeated episodes of microtrauma may accumulate and eventually result in degenerative lesions and clinical tendonitis.

Aging↗
PubMed1997

An age-related study of morphology and cross-link composition of collagen fibrils in the digital flexor tendons of young thoroughbred horses.

The superficial digital flexor tendon is the most commonly injured tendon in the racing Thoroughbred. Despite the clinical significance of this structure, only limited data exist regarding normal age-related morphology of the tensile units, the collagen fibrils. The age at which these collagen fibrils become mature in composition and structure may be of importance. Consequently, the association of age and collagen fibril crosslink composition, diameter distribution and crimp morphology in the superficial and deep digital flexor tendons of Thoroughbreds up to and including three years of age has been studied. Replacement of immature crosslinks, peaking of the collagen fibril mass-average diameter and collagen fibril index, and stabilization of collagen crimp morphology changes supported the hypothesis that both digital flexor tendons become mature in structure by two years of age.

Aging↗
PubMed1996

Heptad breaks in alpha-helical coiled coils: stutters and stammers.

The discontinuities found in heptad repeats of alpha-helical coiled-coil proteins have been characterized. A survey of 40 alpha-fibrous proteins reveals that only two classes of heptad breaks are prevalent: the stutter, corresponding to a deletion of three residues, and the newly identified "stammer," corresponding to a deletion of four residues. This restriction on the variety of insertions/deletions encountered gives support to a unifying structural model, where different degrees of supercoiling accommodate the observed breaks. Stutters in the hemagglutinin coiled-coil region have previously been shown to produce an underwinding of the supercoil, and we show here how, in other cases, stammers would lead to overwinding. An analysis of main-chain structure also indicates that the mannose-binding protein, as well as hemagglutinin, contains an underwound coiled-coil region. In contrast to knobs-into-holes packing, these models give rise to non-close-packed cores at the sites of the heptad phase shifts. We suggest that such non-close-packed cores may function to terminate certain coiled-coil regions, and may also account for the flexibility observed in such long alpha-fibrous molecules as myosin. The local underwinding or overwinding caused by these specific breaks in the heptad repeat has a global effect on the structure and can modify both the assembly of the protein and its interaction properties.

Protein Conformation↗
PubMed1996

Hard alpha-keratin intermediate filaments: an alternative interpretation of the low-angle equatorial X-ray diffraction pattern, and the axial disposition of putative disulphide bonds in the intra- and inter-protofilamentous networks.

STEM and electron microscopy have indicated that intermediate filaments (IF) may contain protofilaments and protofibrils, possibly with four and eight chains in section, respectively. Two important questions related to this observation but pertaining specifically to the structure of the hard alpha-keratin IF have been addressed in this work. The first relates to the apparently smaller number of chains in section (and hence protofilaments and/or protofibrils) in hard alpha-keratin IF as compared to epidermal keratin, vimentin and neurofilament IF. From the low-angle equatorial X-ray diffraction patterns the observed maxima were interpreted by others as arising from the Fourier transform of a uniformly dense cylinder of diameter 7.45 nm and, by implication, containing about 23 chains in section. Here it is shown that a simple, alternative density distribution for the IF is compatible with both the positions of the observed equatorial diffraction maxima and with 32 chains in section, thus allowing the possibility that IF from diverse sources (hard alpha-keratin, epidermal keratin, vimentin and neurofilaments) have a common number of chains in section and hence the same number of subfilamentous elements. The second point relates to the stabilisation of the protofilaments and/or protofibrils. It has been shown that the relative axial alignment of molecules in hard alpha-keratin IF is similar but not identical to that in both epidermal keratin and vimentin IF. A key difference involves the presence of a small gap (about 1.3-1.4nm) between similarly directed molecules as distinct from the head-to-tail overlap between parallel molecules that appears to be a characteristic of all other IF, and which is believed to have a major role in stabilising these assemblies. In contrast, it was proposed that the hard alpha-keratin IF gains its stability in part through a net work of intermolecular disulphide bonds. Here it is shown that disulphide bonds may exist between spatially-adjacent molecules one, two, three or four apart (and beyond) in the proposed surface lattice structure. This indicates the possibility that subfibrillar structural elements consisting of discrete groupings of disulphide-bonded molecules may be present in hard alpha-keratin IF.

Intermediate Filaments↗
PubMed1996

The molecular structure of reptilian keratin.

X-ray diffraction and electron microscope studies of hard keratins (e.g. claws, scales, feathers and hair) have shown that they all have a filamentous texture but that the molecular structure of the filaments in mammalian keratins is quite different from that in avian keratins. The framework of the filaments in mammalian keratin consists of two-strand coiled coils of alpha-helices whereas the framework in avian keratins is composed of beta-sheets. Reptilian hard keratins have not been studied in detail but the X-ray diffraction pattern is very similar to that obtained from avian hard keratins leading to the supposition that the framework of the filaments is also composed of beta-sheets. The present contribution describes an analysis of the sequence of a lizard claw protein using structural probes which reveal the origins of the common structural features of the filaments in avian and reptilian keratin.

Animals↗
PubMed1996

Cloning of novel kinectin splice variants with alternative C-termini: structure, distribution and evolution of mouse kinectin.

The analysis of cDNA clones encoding novel variant forms of mouse kinectin, an endoplasmic reticulum (ER)-bound receptor for the motor protein kinesin, is reported. Kinesin and cytoplasmic dynein are involved in mediating the anterograde and retrograde movements of intracellular vesicles along the microtubule network. The amino acid sequence deduced from kinectin cDNA isolated from mouse spleen cell and testis libraries revealed a long signal peptide or transmembrane sequence, and a 328 amino acid residue globular N-terminal domain adjacent to a much larger 858-999-residue C-terminal coiled-coil rod domain. The C-terminal domain was composed of 18 coiled-coil regions formed from multiple contiguous heptad repeats which undergo alternative splicing as evidenced by the presence of at least five small (23-33 amino acid residue) insertion sequences scattered throughout. The inserts are present in any one of a number of combinations, generating an array of novel kinectin variants. Insert 5 contains a termination codon, producing a C-terminus that is highly homologous to that of human kinectin. Three out of five mouse kinectin clones lack insert 5, generating a novel eleven amino acid C-terminus encoded by sequence that extends past the insertion site. The existence of alternative C-termini may have functional relevance given that the C-termini are exposed for interaction with kinesin, whereas the globular N-terminus is embedded in the ER membrane. Alternative C-termini represent candidate modifications that could determine specificity of binding to kinesin or cytoplasmic dynein, and the switching of directionality of movement. The cDNA hybridized to 4.5 kb transcripts expressed in all mouse cell lines and tissues examined, which provides the first indication that the kinectins are very widely distributed. Mouse kinectin is 42% similar over a 203 amino acid region to the chicken extracellular cardiac morphogen ES/130, whose canine homologue containing an inserted sequence of 10 amino acids repeated 54 times in tandem, is a ribosome receptor expressed on the ER. Mouse kinectin shares 64 and 83% identity, respectively, with its M(r) 160000 chicken and human kinectin homologues. There is a two-fold molar excess of kinectin over kinesin in unextracted vesicles, suggesting that kinectin might be a dimer. The electrostatic properties of the coiled-coil region of mouse kinectin, together with the relative frequencies of residues in particular positions within the heptad repeats support this notion.

Alternative Splicing↗
PubMed1996

Clinical and histologic observations of sites implanted with intraoral autologous bone grafts or allografts. 15 human case reports.

The cases reported in this paper were treated at 7 different clinical centers and present clinical and histologic observations from 15 patients and 21 human biopsies. The biopsies were taken from extraction sockets or dental implant sites which were grafted with either autologous intra-oral bone (6 sites), demineralized freeze-dried bone (DFDBA) (7 sites), or mineralized freeze-dried bone (MFDBA) (7 sites), or a combination of autologous bone, DFDBA and a barrier membrane (1 site). Six sites were grafted with DFDBA and augmented with expanded polytetrafluoroethylene (ePTFE) barrier membranes. Biopsies for histological evaluation were taken 4 to 13 months after implantation. A bone scoring system of 0 to 4 was used to evaluate the sections for dead implanted particles or the presence of vital bone. A bone score of 3 indicated the presence of dead implant material, blood vessels, islands of cartilage, osteoblasts, and new bone formation. A score of 4 indicated total replacement of the implanted material by the host bone. The average bone score for sites which received autologous bone was 2.33; for DFDBA sites, 0.98; and MFDBA was 0.18. The over-riding histologic characteristic of sites implanted with DFDBA or MFDBA was retention of non-vital graft particles within fibrous connective tissue. Biopsies taken adjacent to the host bed demonstrated incorporation of the allografts (osteoconduction). Sites grafted with autologous bone chips also demonstrated non-vital bone chips surrounded by vital host bone (osteoconduction). Sites which received barrier membranes did not appear to improve or impair bone healing of the augmented sites. Autologous bone chips harvested from within the oral cavity as well as allografts may serve as biologic fillers, but do not apparently contribute to osteoinduction. Autologous bone will eventually be resorbed and replaced by the host. DFDBA and MFDBA are resorbed very slowly and apparently do not contribute to osteoinduction. Allografts apparently are not resorbed by osteoclasts and therefore their continued use around dental implants is questioned.

Adult↗