COVALENT STRUCTURE OF COLLAGEN - AMINO-ACID-SEQUENCE OF ALPHA-1(III)-CB5 FROM TYPE-III COLLAGEN OF HUMAN-LIVER
COVALENT STRUCTURE OF COLLAGEN - AMINO-ACID-SEQUENCE OF ALPHA-1(III)-CB5 FROM TYPE-III COLLAGEN OF HUMAN-LIVER
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DOI：
10.1021/bi00549a008
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发表时间：
1980-01-01
期刊：
影响因子：
2.9
通讯作者：
KANG, AH
中科院分区：
文献类型：
--
作者：
SEYER, JM;MAINARDI, C;KANG, AH
以人肝为原料，采用有限胃酶消化、差示盐析、羧甲基纤维素层析等方法制备了III型胶原蛋白。经CNBR消化，得到10个不同的多肽。用羧甲基纤维素层析法进一步纯化α1(III)-CB5多肽，并测定其氨基酸序列。使用完整的α1(III)-CB5、胰酶和热裂解肽以及羟胺衍生片段的自动Edman降解来建立总序列。用纯化的类风湿滑膜胶原酶消化天然III型胶原，确定了α1(III)-CB5序列中包含的哺乳动物胶原酶位置。胶原酶的裂解发生在单一的Gly.sbd.Ile键上，1个三联体在相应的I型胶原的特定裂解位点之前。人肝脏III型胶原的序列包括α1(III)-CB3-7-6-1-8-10-2-4-5。这些对应于α1(I)-CB0-1-2-4-5-8-3-7残基11-804的同源区。
Type III collagen was prepared from human liver by limited pepsin digestion, differential salt precipitation and carboxymethylcellulose chromatography. Ten distinct peptides were obtained by CNBr digestion. The peptide .alpha.1(III)-CB5 was further purified by carboxymethylcellulose chromatography, and its amino acid sequence was determined. Automatic Edman degradation of intact .alpha.1(III)-CB5, tryptic and thermolytic peptides and hydroxylamine-derived fragments was used to establish the total sequence. The mammalian collagenase site contained in the .alpha.1(III)-CB5 sequence was ascertained by digestion of native type III collagen with purified rheumatoid synovial collagenase. Collagenase cleavage occurred at a single Gly.sbd.Ile bond, 1 triplet before the corresponding specific cleavage site of type I collagen. The sequence of human liver type III collagen includes .alpha.1(III)-CB3-7-6-1-8-10-2-4-5. These correspond to the homologous region of .alpha.1(I)-CB0-1-2-4-5-8-3-7 residues 11-804.