A Type VI Collagen-related Glycopolypeptide is the Major Concanavalin A-binding Component in Pig Skin

Overview

Journal Biochem J

Specialty Biochemistry

Date 1989 Jan 1

PMID 2920030

Citations 2

Authors

I A King

A Tabiowo

P R Fryer

F M Pope

Affiliations

Soon will be listed here.

Abstract

The major concanavalin A-binding component in urea/deoxycholate/mercaptoethanol extracts of pig skin was a collagenous disulphide-cross-linked glycopolypeptide with an apparent molecular mass of 150 kDa and a pI of 5.5. Antiserum against the electrophoretically purified glycopolypeptide gave strong dermal staining similar to that seen with fluorescent concanavalin A. Immunocytochemical labelling showed prominent labelling of 3-4 nm dermal microfilaments, particularly those associated with dermal blood vessels and mast cells. Immunoblotting with authentic antiserum indicated that the major skin glycopolypeptide was probably identical with collagen-like glycoprotein, the tissue form of the alpha 1/alpha 2 subunits of type VI collagen. This was confirmed by immunoblotting of authentic type VI collagen from pepsin-treated pig skin. Immunoblotting, metabolic labelling with [3H]glucosamine and immune precipitation showed that an immunoreactive collagenous glycopolypeptide was synthesized and secreted by cultured pig skin fibroblasts. The results suggest that type VI collagen is the major concanavalin A-binding component in pig skin.

Citing Articles

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Molecular composition of type VI collagen. Evidence for chain heterogeneity in mammalian tissues and cultured cells.

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References

Gibson M, Cleary E . A collagen-like glycoprotein from elastin-rich tissues. Biochem Biophys Res Commun. 1982; 105(4):1288-95. DOI: 10.1016/0006-291x(82)90926-3. View

King I, Tabiowo A, Fryer P . Evidence that major 78-44-kD concanavalin A-binding glycopolypeptides in pig epidermis arise from the degradation of desmosomal glycoproteins during terminal differentiation. J Cell Biol. 1987; 105(6 Pt 2):3053-63. PMC: 2114695. DOI: 10.1083/jcb.105.6.3053. View

Zieske J, BERNSTEIN I . Modification of cell surface glycoprotein: addition of fucosyl residues during epidermal differentiation. J Cell Biol. 1982; 95(2 Pt 1):626-31. PMC: 2112967. DOI: 10.1083/jcb.95.2.626. View

Lehto V, Virtanen I . Immunolocalization of a novel, cytoskeleton-associated polypeptide of Mr 230,000 daltons (p230). J Cell Biol. 1983; 96(3):703-16. PMC: 2112408. DOI: 10.1083/jcb.96.3.703. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Bonner W, Laskey R . A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974; 46(1):83-8. DOI: 10.1111/j.1432-1033.1974.tb03599.x. View

Ofarrell P . High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975; 250(10):4007-21. PMC: 2874754. View

Chung E, Rhodes K, Miller E . Isolation of three collagenous components of probable basement membrane origin from several tissues. Biochem Biophys Res Commun. 1976; 71(4):1167-74. DOI: 10.1016/0006-291x(76)90776-2. View

Holt P, ANGLIN Jr J, Nordquist R . Localization of specific carbohydrate configurations in human skin using fluorescein-labelled lectins. Br J Dermatol. 1979; 100(3):237-45. DOI: 10.1111/j.1365-2133.1979.tb06194.x. View

10.

Towbin H, Staehelin T, Gordon J . Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979; 76(9):4350-4. PMC: 411572. DOI: 10.1073/pnas.76.9.4350. View

11.

Furuto D, Miller E . Isolation of a unique collagenous fraction from limited pepsin digests of human placental tissue. Characterization of one of the constituent polypeptide chains. J Biol Chem. 1980; 255(1):290-5. View

12.

Brabec R, Peters B, BERNSTEIN I, Gray R, Goldstein I . Differential lectin binding to cellular membranes in the epidermis of the newborn rat. Proc Natl Acad Sci U S A. 1980; 77(1):477-9. PMC: 348294. DOI: 10.1073/pnas.77.1.477. View

13.

Jander R, Rauterberg J, Voss B, von Bassewitz D . A cysteine-rich collagenous protein from bovine placenta. Isolation of its constituent polypeptide chains and some properties of the non-denatured protein. Eur J Biochem. 1981; 114(1):17-25. View

14.

Furuto D, Miller E . Characterization of a unique collagenous fraction from limited pepsin digests of human placental tissue: molecular organization of the native aggregate. Biochemistry. 1981; 20(6):1635-40. DOI: 10.1021/bi00509a035. View

15.

Fryer P, Wells C, Ratcliffe A . Technical difficulties overcome in the use of Lowicryl 4KM EM embedding resin. Histochemistry. 1983; 77(1):141-3. DOI: 10.1007/BF00496645. View

16.

Nemanic M, WHITEHEAD J, Elias P . Alterations in membrane sugars during epidermal differentiation: visualization with lectins and role of glycosidases. J Histochem Cytochem. 1983; 31(7):887-97. DOI: 10.1177/31.7.6854004. View

17.

Odermatt E, Risteli J, van Delden V, Timpl R . Structural diversity and domain composition of a unique collagenous fragment (intima collagen) obtained from human placenta. Biochem J. 1983; 211(2):295-302. PMC: 1154359. DOI: 10.1042/bj2110295. View

18.

Gibson M, Cleary E . Distribution of CL glycoprotein in tissues: an immunohistochemical study. Coll Relat Res. 1983; 3(6):469-88. DOI: 10.1016/s0174-173x(83)80027-2. View

19.

Hessle H, Engvall E . Type VI collagen. Studies on its localization, structure, and biosynthetic form with monoclonal antibodies. J Biol Chem. 1984; 259(6):3955-61. View

20.

Carter W . The role of intermolecular disulfide bonding in deposition of GP140 in the extracellular matrix. J Cell Biol. 1984; 99(1 Pt 1):105-14. PMC: 2275619. DOI: 10.1083/jcb.99.1.105. View