Wohllebe M, Carmichael D J
Biochem J. 1979 Sep 1;181(3):667-76. doi: 10.1042/bj1810667.
alpha- and beta-Chains were isolated by sequential ion-exchange and gel-filtration chromatography of guanidinium chloride-soluble dentine collagen obtained from Tris/NaCl-extracted EDTA-demineralized lathyritic-rat incisors. The alpha-chains were identified as alpha 1 I and alpha 2 by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and amino acid analysis of the intact chains and their CNBr peptides. The dentine alpha-chains exhibited higher lysine hydroxylation and phosphate content, but lower hydroxylysine glycosylation, than alpha-chains from skin. Increased lysine hydroxylation was observed in the helical sequences. The alpha 1 I/alpha 2 ratio was approx. 3:1, and was presumably due to the presence of (alpha 1 I)3 molecules along with (alpha 1 I)2 alpha 2 molecules as shown recently for neutral-salt-soluble dentine collagen [Wohllebe & Carmichael (1978) Eur. J. Biochem. 92, 183--188]. In the borohydride-reduced beta 11- and beta 12-chains from guanidinium chloride-soluble dentine collagen, the reduced cross-links hydroxylysinohydroxynorleucine and hydroxylysinonorleucine were present. A higher proportion of hydroxylysinonorleucine in the reduced beta 12-chain probably reflects differences in extent of hydroxylation of specific lysine residues of the alpha 1 I- and alpha 2-chains.
通过对从经Tris/NaCl提取、EDTA脱矿的致跛行大鼠切牙中获得的氯化胍可溶性牙本质胶原进行连续离子交换和凝胶过滤色谱法，分离出α链和β链。通过十二烷基硫酸钠/聚丙烯酰胺凝胶电泳以及完整链及其CNBr肽的氨基酸分析，将α链鉴定为α1I和α2。与皮肤来源的α链相比，牙本质α链表现出更高的赖氨酸羟基化程度和磷酸盐含量，但羟基赖氨酸糖基化程度较低。在螺旋序列中观察到赖氨酸羟基化增加。α1I/α2的比例约为3:1，推测这是由于存在(α1I)3分子以及(α1I)2α2分子，正如最近对中性盐可溶性牙本质胶原所显示的那样[Wohllebe & Carmichael (1978) Eur. J. Biochem. 92, 183 - 188]。在来自氯化胍可溶性牙本质胶原的硼氢化物还原的β11和β12链中，存在还原的交联物羟赖氨酰羟正亮氨酸和羟赖氨酰正亮氨酸。还原的β12链中较高比例的羟赖氨酰正亮氨酸可能反映了α1I链和α2链特定赖氨酸残基羟基化程度的差异。
