Abstract
Immunological studies of rat skin collagen were carried out with a sensitive and quantitative radioimmunoassay. Hyperimmune rabbit antisera to rat skin collagen and isolated α2 chains were used. Iodine-labeled α chains and CNBr-produced peptides served as test antigens, and native collagen, α chains, and CNBr peptides were employed as inhibitors in the assay. The α1 and α2 chains were immunologically distinct. Although the α1 chain was not immunogenic, antibodies to α1 were detected in antisera to the intact collagen molecule. The major antigenic determinant of the α1 chain was located in α1-CB6 which constitutes the carboxy-terminal region of the chain. The α2 chain contained two non-cross-reacting antigenic determinants, one in the amino-terminal region (α2-CB1) and the other in the carboxy-terminal region (α2-CB5) of the chain. The native collagen molecule was less effective than isolated α chains in inhibiting binding of labeled peptides to antisera, indicating that antigenic determinants were less accessible in the triple helical molecule. These immunologic studies are consistent with preliminary comparative biochemical data which indicate that interspecies structural differences in collagen predominate at both the amino- and carboxy-terminal ends of the chains.
Full Text
The Full Text of this article is available as a PDF (838.2 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- ARNON R., SELA M. Studies on the chemical basis of the antigenicity of proteins. 2. Antigenic specificity of polytyrosyl gelatins. Biochem J. 1960 Apr;75:103–109. doi: 10.1042/bj0750103. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bensusan H. B. Protein crosslinkages. I. Effect of iodination on the time-dependent solubility changes of reconstituted collagen fibrils. Arch Biochem Biophys. 1966 Jul;115(1):77–83. doi: 10.1016/s0003-9861(66)81040-8. [DOI] [PubMed] [Google Scholar]
- Bornstein P. Collagen: relatively invariant (helical) and variable (nonhelical) regions. Science. 1968 Aug 9;161(3841):592–593. doi: 10.1126/science.161.3841.592. [DOI] [PubMed] [Google Scholar]
- Bornstein P., Nesse R. The comparative biochemistry of collagen: the structure of rabbit skin colllagen and its relevance to immunochemical studies of collagen. Arch Biochem Biophys. 1970 Jun;138(2):443–450. doi: 10.1016/0003-9861(70)90367-x. [DOI] [PubMed] [Google Scholar]
- Bornstein P., Piez K. A. The nature of the intramolecular cross-links in collagen. The separation and characterization of peptides from the cross-link region of rat skin collagen. Biochemistry. 1966 Nov;5(11):3460–3473. doi: 10.1021/bi00875a012. [DOI] [PubMed] [Google Scholar]
- Butler W. T., Piez K. A., Bornstein P. Isolation and characterization of the cyanogen bromide peptides from the alpha-1 chain of rat skin collagen. Biochemistry. 1967 Dec;6(12):3771–3780. doi: 10.1021/bi00864a022. [DOI] [PubMed] [Google Scholar]
- Click E. M., Bornstein P. Isolation and characterization of the cyanogen bromide peptides from the alpha 1 and alpha 2 chains of human skin collagen. Biochemistry. 1970 Nov 24;9(24):4699–4706. doi: 10.1021/bi00826a012. [DOI] [PubMed] [Google Scholar]
- Davison P. F., Levine L., Drake M. P., Rubin A., Bump S. The serologic specificity of tropocollagen telopeptides. J Exp Med. 1967 Aug 1;126(2):331–346. doi: 10.1084/jem.126.2.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fietzek P. P., Piez K. A. Isolation and characterization of the cyanogen bromide peptides from the alpha 2 chain of rat skin collagen. Biochemistry. 1969 May;8(5):2129–2133. doi: 10.1021/bi00833a052. [DOI] [PubMed] [Google Scholar]
- Helmkamp R. W., Contreras M. A., Bale W. F. I-131-labeling of proteins by the iodine monochloride method. Int J Appl Radiat Isot. 1967 Nov;18(11):737–746. doi: 10.1016/0020-708x(67)90011-7. [DOI] [PubMed] [Google Scholar]
- Kang A. H., Bornstein P., Piez K. A. The amino acid sequence of peptides from the cross-linking region of rat skin collagen. Biochemistry. 1967 Mar;6(3):788–795. doi: 10.1021/bi00855a019. [DOI] [PubMed] [Google Scholar]
- Kirrane J. A., Robertson W. V. The antigenicity of native and tyrosylated neutral-salt-soluble rat collagen. Immunology. 1968 Jan;14(1):139–148. [PMC free article] [PubMed] [Google Scholar]
- Kivirikko K. I., Laitinen O., Prockop D. J. Modifications of a specific assay for hydroxyproline in urine. Anal Biochem. 1967 May;19(2):249–255. doi: 10.1016/0003-2697(67)90160-1. [DOI] [PubMed] [Google Scholar]
- LeRoy E. C. Precipitating and complement-fixing antibodies to collagen with species and collagen subunit specificity. Proc Soc Exp Biol Med. 1968 Jun;128(2):341–347. doi: 10.3181/00379727-128-33010. [DOI] [PubMed] [Google Scholar]
- Michaeli D., Martin G. R., Kettman J., Benjamini E., Leung D. Y., Blatt B. A. Localization of antigenic determinants in the polypeptide chains of collagen. Science. 1969 Dec 19;166(3912):1522–1524. doi: 10.1126/science.166.3912.1522. [DOI] [PubMed] [Google Scholar]
- Piez K. A., Bladen H. A., Lane J. M., Miller E. J., Bornstein P., Butler W. T., Kang A. H. Comparative studies on the chemistry of collagen utilizing cyanogen bromide cleavage. Brookhaven Symp Biol. 1968 Jun;21(2):345–357. [PubMed] [Google Scholar]
- Piez K. A., Miller E. J., Lane J. M., Butler W. T. The order of the CNBr peptides from the alpha-1 chain of collagen. Biochem Biophys Res Commun. 1969 Nov 20;37(5):801–805. doi: 10.1016/0006-291x(69)90962-0. [DOI] [PubMed] [Google Scholar]
- Pontz B., Meigel W., Rauterberg J., Kühn K. Localization of two species specific antigenic determinants on the peptide chains of calf skin collagen. Eur J Biochem. 1970 Sep;16(1):50–54. doi: 10.1111/j.1432-1033.1970.tb01051.x. [DOI] [PubMed] [Google Scholar]
- Porath J., Axen R., Ernback S. Chemical coupling of proteins to agarose. Nature. 1967 Sep 30;215(5109):1491–1492. doi: 10.1038/2151491a0. [DOI] [PubMed] [Google Scholar]
- ROBBINS J. B., KENNY K., SUTER E. THE ISOLATION AND BIOLOGICAL ACTIVITIES OF RABBIT GAMMA M- AND GAMMA G-ANTI-SALMONELLA TYPHIMURIUM ANTIBODIES. J Exp Med. 1965 Aug 1;122:385–402. doi: 10.1084/jem.122.2.385. [DOI] [PMC free article] [PubMed] [Google Scholar]
- ROTHBARD S., WATSON R. F. Antigenicity of rat collagen. Demonstration of antibody to rat collagen in the renal glomeruli of rats by fluorescence microscopy. J Exp Med. 1961 Jun 1;113:1041–1051. doi: 10.1084/jem.113.6.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]
- ROTHBARD S., WATSON R. F. Antigenicity of rat collagen; reverse anaphylaxis induced in rats by anti-rat collagen serum. J Exp Med. 1956 Jan 1;103(1):57–72. doi: 10.1084/jem.103.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rothbard S., Watson R. F. Immunologic relations among various animal collagens. J Exp Med. 1965 Sep 1;122(3):441–454. doi: 10.1084/jem.122.3.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SCHMITT F. O., LEVINE L., DRAKE M. P., RUBIN A. L., PFAHL D., DAVISON P. F. THE ANTIGENICITY OF TROPOCOLLAGEN. Proc Natl Acad Sci U S A. 1964 Mar;51:493–497. doi: 10.1073/pnas.51.3.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
- STEFFEN C., TIMPL R., WOLFF I. IMMUNOGENICITY AND SPECIFICITY OF COLLAGEN. II. INVESTIGATIONS ABOUT SPECIFICITY OF COLLAGEN AND ITS DERIVATIVES BY HEMAGGLUTINATION AND HEMAGGLUTINATION-INHIBITION OF ANTI-COLLAGEN AND ANTI-PARENT GELATINE IMMUNE SERA. J Immunol. 1964 Oct;93:656–667. [PubMed] [Google Scholar]
- Steffen C., Timpl R. Immunogenicity and specificity of collagen. X. Investigations on collagen determinants by inhibition experiments with peptides obtained after multiple degradation. Z Immunitatsforsch Allerg Klin Immunol. 1970 Jun;139(5):455–467. [PubMed] [Google Scholar]
- Steffen C., Timpl R., Wolff I. Immunogenicity and specificity of collagen. V. Demonstration of three different antigenic determinants on calf collagen. Immunology. 1968 Jul;15(1):135–144. [PMC free article] [PubMed] [Google Scholar]
- Steffen C., Timpl R., Wolff I. Immunogenität und Spezifität von Kollagen. 3. Erzeugung und Nachweis von Anti-Kollagen-Antikörpern und serologischer Vergleich von nativem und denaturiertem löslichem Kollagen. Z Immunitatsforsch Allerg Klin Immunol. 1967 Nov;134(1):91–107. [PubMed] [Google Scholar]
- Timpl R., Wolff I., Furthmayr H., Steffen C. Immunogenicity and specificity of collagen. VI. Separation of antibody fractions with restricted specificity from anti-collagen sera using an immunoadsorbent technique. Immunology. 1968 Jul;15(1):145–151. [PMC free article] [PubMed] [Google Scholar]
- Vuust J., Lane J. M., Fietzek P. P., Miller E. J., Piez K. A. The order of the CNBr peptides from the alpha 2 chain of collagen. Biochem Biophys Res Commun. 1970 Feb 20;38(4):703–708. doi: 10.1016/0006-291x(70)90638-8. [DOI] [PubMed] [Google Scholar]
- von der Mark K., Rexrodt F., Wendt P., Kühn K. Isolation and characterization of cyanogen bromide-cleaved peptides from a collagenase digest of alpha 1-chain of calf skin collagen. Eur J Biochem. 1970 Sep;16(1):143–151. doi: 10.1111/j.1432-1033.1970.tb01065.x. [DOI] [PubMed] [Google Scholar]