LOCATION OF THE INTERMOLECULAR CROSS-LINKS IN BOVINE DENTIN COLLAGEN, SOLUBILIZATION WITH TRYPSIN AND ISOLATION OF CROSS-LINK PEPTIDES CONTAINING DIHYDROXYLYSINONORLEUCINE AND PYRIDINOLINE
LOCATION OF THE INTERMOLECULAR CROSS-LINKS IN BOVINE DENTIN COLLAGEN, SOLUBILIZATION WITH TRYPSIN AND ISOLATION OF CROSS-LINK PEPTIDES CONTAINING DIHYDROXYLYSINONORLEUCINE AND PYRIDINOLINE
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DOI：
10.1016/0006-291x(81)91497-2
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发表时间：
1981-01-01
影响因子：
3.1
通讯作者：
MECHANIC, GL
中科院分区：
文献类型：
--
作者：
KUBOKI, Y;TSUZAKI, M;MECHANIC, GL
[3H]NaBH4还原后的牛牙本质胶原在60℃下变性。C反应1小时，然后用胰蛋白酶消化。消化液基本上仍是不溶性悬浮液，但如果在60度再次加热，发现99%的牙本质胶原蛋白可以被溶解。C敷15分钟。通过Sephadex G50、phosphocellulose和DEAE-cellulose色谱柱的顺序层析，分离得到两个交联的胰蛋白酶肽。分离得到的一个肽段为59个残基交联肽段，包含1个二羟基赖氨酸残基；另一个肽段为103个残基，包含1个吡啶碱残基。氨基酸组成与鉴定的第59个残基肽序列一致。1-CB4-5（76-90）与。alpha序列相连。1-CB6 （990-23c）和103残基肽作为序列76-90连接到序列990-23c的2。这些结果有力地支持了先前提出的二羟基赖氨酸氨基亮氨酸和吡啶啉之间的前体产物关系。
[3H]NaBH4 reduced bovine dentin collagen was denatured at 60.degree. C for 1 h and then digested with trypsin. The digest was still substantially insoluble suspension, but it was found that 99% of dentin collagen can be solubilized if the digest was heated again at 60.degree. C for 15 min. Two cross-linked tryptic peptides were isolated from this digest by sequential chromatographies on Sephadex G50, phosphocellulose and DEAE-cellulose column. One isolated peptide was characterized as a 59 residue cross-linked peptide including 1 residue of dihydroxylysinonorleucine and the other was 103 residue including 1 residue of pyridinoline. The amino acid compositions were consistent with the identification of the 59 residue peptide as the sequence in .alpha.1-CB4-5 (76-90) linked to the sequence of .alpha.1-CB6 (990-23c), and the 103 residue peptide as the sequence 76-90 linked to 2 of the sequence 990-23c. These results strongly support the previously proposed precursor-product relationship between dihydroxylysinonorleucine and pyridinoline.