Physical Evidence for the Asembly of A and B Chains of Human Placental Collagen in a Single Triple Helix

Bentz, Hanne; Bächinger, Hans Peter; Glanville, Robert W.; Kühn, Klaus

doi:10.1111/j.1432-1033.1978.tb12778.x

Cited by 161 publications

(58 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3). However, previous in-vitro data have demonstrated that a [a2(V)I3 homotrimer is unstable (Bentz et al 1978). In addition, no single a2(V) collagen chain can be present in the pepsinsolubilized matrix material since it is resistant to pepsin digestion.…”

Section: Discussionmentioning

confidence: 94%

The human rhabdomyosarcoma cell line A204 lays down a highly insoluble matrix composed mainly of α1 type‐XI and α2 type‐V collagen chains

Kleman

Hartmann

Ramirez³

et al. 1992

European Journal of Biochemistry

View full text Add to dashboard Cite

The biosynthesis of collagen by the A204 cell line was examined using polyclonal antibodies raised against collagen typeV and type XI. The study of the pepsin-digested collagen showed that it is composed mainly of al(X1) and a2(V) collagen chains in an apparent 2 : l ratio, suggesting the formation of heterotypic molecules [al(XI)lza2(V). The existence of this chain stoichiometry was further demonstrated by immunoprecipitation of the molecule with an antibody recognizing a2(V) but not al(X1) collagen chains.Electron microscopy analyses of 24-h cultures showed that this matrix is composed of thin fibrils, that can be decorated with immunogold-labelled anti-(type-V collagen) IgG, but not with anti-(type-XI collagen) IgG. The collagen matrix laid down by A204 cells is highly insoluble. In the presence of P-aminopropionitrile, an inhibitor of lysyl oxidase, only a small proportion of intact collagen could be extracted without proteolytic treatment. Immunoblotting of intact medium collagen from cultures performed in the presence of P-aminopropionitrile showed four distinct bands with each antibody. The migration of the bands, stained with anti-(type-V collagen) IgG, had apparent molecular masses of 127, 149, 161 and 198 kDa (compared to globular standards) while the bands stained with anti-(type-XI collagen) IgG had apparent masses of 145, 182, 207 and 225 kDa.These data indicate that type-V and type-XI collagen chains can assemble in heterotypic isoforms. In this system, the synthesized isoforms are able to aggregate into a highly cohesive matrix and they undergo a proteolytic processing closely similar to that of other fibrillar collagens.Collagen types V and XI are members of the collagens forming quarter-staggered fibrils (Woodbury et al., 1989;Kimura et al., 1989;Weil et al., 1987). Type-V collagen is commonly found as a quantitatively minor component in collagen-type-I-containing extracellular matrices such as those of placenta, skin and bone (Niyibizi et al., 1984;Birk et al., 1988;Niyibizi and Eyre, 1989a). Type-XI collagen is found in association with type I1 in cartilaginous matrices (Mendler et al., 1989). This led to the dichotomous view that fibrilforming collagens can be subdivided into two sub-groups, according to their tissue distribution, (a) collagens associated with collagen type I in non-cartilaginous tissues, in particular type V, (b) collagens associated with type I1 in cartilaginous matrices, such as type-XI collagen. Recent data indicate that collagen types V and XI are not so strictly partitioned as initially thought. First, they are thought to play a similar role in collagen fibrillogenesis (Morris and Bichinger, 1987;Morris et al., 1990), possibly by forming a core to the fibril (Birk et al., 1990;Fessler et al., 1985a;Mendler et al., 1989). These two molecules are both immunologically masked and require partial collagen fibril disruption for immunological detection (Birk et al., 1988). Furthermore, close homologies have been reported between type-V and type-XI collagen chains. The al(X1) a...

show abstract

Section: Discussionmentioning

confidence: 94%

The human rhabdomyosarcoma cell line A204 lays down a highly insoluble matrix composed mainly of α1 type‐XI and α2 type‐V collagen chains

Kleman

Hartmann

Ramirez³

et al. 1992

European Journal of Biochemistry

View full text Add to dashboard Cite

show abstract

“…However, Col5a1 À/À embryos die at approximately 10.5 dpc, 9 2 days earlier in gestation than the Col5a2 À/À embryos described here. Because a2(V) chains are thought to be unstable in the absence of a1(V) chains with which to combine, 8,28 the phenotype of Col5a1 À/À embryos likely results from a complete absence of both a1(V) and a2(V) chains and thus a lack of collagen V. However, in the absence of a2(V) chains with which to combine, a1(V) chains form stable a1(V) 3 homotrimers. 28,29 Thus, the 2 additional days of embryonic survival of Col5a2 À/À , compared with Col5a1 À/À embryos, suggests that a1(V) 3 homotrimers can functionally compensate, in part, for the loss of a1(V) 2 a2(V) heterotrimers.…”

Section: Discussionmentioning

confidence: 99%

“…Because a2(V) chains do not appear to be capable of forming stable a2(V) homotrimers in the absence of a1(V) chains with which to combine, 8,28 the phenotype of Col5a1 À/À embryos is thought to be associated with total absence of both a1(V) and a2(V) chains because unstable, nontrimeric a2(V) chains would be degraded rather than incorporated into the ECM in Col5a1 À/À tissues. In contrast, the phenotype of Col5a2 À/À embryos should be associated not only with the absence of normal a1(V) 2 a2(V) heterotrimers but also with the presence of aberrant a1(V) 3 homotrimers because a1(V) chains in the absence of a2(V) chains form stable a1(V) 3 homotrimers.…”

Section: A1(v) Collagen Chains Associate With Collagen I Fibrils In Tmentioning

confidence: 99%

Homozygosity and Heterozygosity for Null Col5a2 Alleles Produce Embryonic Lethality and a Novel Classic Ehlers-Danlos Syndrome–Related Phenotype

Park

Phillips

Pfeiffer

et al. 2015

The American Journal of Pathology

View full text Add to dashboard Cite

Null alleles for the COL5A1 gene and missense mutations for COL5A1 or the COL5A2 gene underlie cases of classic Ehlers-Danlos syndrome, characterized by fragile, hyperextensible skin and hypermobile joints. However, no classic Ehlers-Danlos syndrome case has yet been associated with COL5A2 null alleles, and phenotypes that might result from such alleles are unknown. We describe mice with null alleles for the Col5a2. Col5a2 À/À homozygosity is embryonic lethal at approximately 12 days post conception. Unlike previously described mice null for Col5a1, which die at 10.5 days post conception and virtually lack collagen fibrils, Col5a2 À/À embryos have readily detectable collagen fibrils, thicker than in wild-type controls. Differences in Col5a2 À/À and Col5a1 À/À fibril formation and embryonic survival suggest that a1(V) 3 homotrimers, a rare collagen V isoform that occurs in the absence of sufficient levels of a2(V) chains, serve functional roles that partially compensate for loss of the most common collagen V isoform. Col5a2 þ/À adults have skin with marked hyperextensibility and reduced tensile strength at high strain but not at low strain. Col5a2 þ/À adults also have aortas with increased compliance and reduced tensile strength. Results thus suggest that COL5A2 þ/À humans, although unlikely to present with frank classic Ehlers-Danlos syndrome, are likely to have fragile connective tissues with increased susceptibility to trauma and certain chronic pathologic conditions. Collagen V is a low-abundance fibrillar collagen widely distributed in vertebrate tissues as a1(V) 2 a2(V) heterotrimers, 1 which are incorporated into growing fibrils with the more abundant collagen I and involved in regulating the geometry and tensile strength of the resulting collagen I/V heterotypic fibrils. 2,3 Mutations in the genes encoding either the a1(V) 4 or a2(V) 5 chain can result in the human heritable connective tissue disorder classic Ehlers-Danlos syndrome (cEDS), clinical hallmarks of which include skin hyperextensibility, atrophic scarring, and joint hypermobility, with patients also often presenting with easy bruising and bleeding. 6 At the molecular level, the collagen fibrils of cEDS skin have variability in diameter not seen in normal skin and include large diameter collagen fibril aggregates with abnormal cauliflower-like shapes when viewed in cross section. 6 Deficits in the tensile strength of cEDS collagen fibrils are inferred from the hyperextensibility and fragility of cEDS skin and the hypermobility of cEDS joints.Most cEDS cases that have been characterized at the molecular level are heterozygous for null alleles of the a1(V) chain gene COL5A1, 7 resulting in the deposition of haploinsufficient levels of normal collagen V in tissues, with excess a2(V) chains unable to form stable triple helical molecules or be incorporated into the extracellular matrix (ECM). 8

show abstract

“…Lathyritic chick type I1 collagen and bovine nasal type I1 collagen were generous gifts from Dr. Klaus von der Mark and Dr. Rupert Tirripl, respectively (Max Planck Institut fur Biochemie, Munich, FRG) (16). Type V (AB) collagen was purified from a pepsin digest of human placenta (18). All collagens were lyophilized and then stored and, prior to use, were dissolved in 0.1M acetic acid.…”

Section: Methodsmentioning

confidence: 99%

Characterization of the antibody response in mice with type II collagen–induced arthritis, using monoclonal anti–type II collagen antibodies

Holmdahl

Rubin

Klareskog

et al. 1986

Arthritis & Rheumatism

369

242

View full text Add to dashboard Cite

Twenty monoclonal antibodies reactive with type I1 collagen were characterized as to their determinant specificity and their reactivity with cartilage-derived components. The monoclonal antibodies reacted with 7 different epitopes on the native type I1 collagen triple helical structure. Antibodies defining 3 of these epitopes Wcurred more frequently in sera from arthritic mice than in sera from nonarthritic mice. In vivo injection of some selected autoreactive antibodies caused synovitis, but in no case did it give rise to full-blown arthritis.Autoimmune reactions, particularly formation of autoantibody against cartilage-derived collagenous components, have frequently been reported in connecPresented by Dr. Holmdahl in partial fulfillment of the requirements for

show abstract

Physical Evidence for the Asembly of A and B Chains of Human Placental Collagen in a Single Triple Helix

Cited by 161 publications

References 18 publications

The human rhabdomyosarcoma cell line A204 lays down a highly insoluble matrix composed mainly of α1 type‐XI and α2 type‐V collagen chains

The human rhabdomyosarcoma cell line A204 lays down a highly insoluble matrix composed mainly of α1 type‐XI and α2 type‐V collagen chains

Homozygosity and Heterozygosity for Null Col5a2 Alleles Produce Embryonic Lethality and a Novel Classic Ehlers-Danlos Syndrome–Related Phenotype

Characterization of the antibody response in mice with type II collagen–induced arthritis, using monoclonal anti–type II collagen antibodies

Contact Info

Product

Resources

About