EVIDENCE FOR GLUCOSE-MEDIATED COVALENT CROSS-LINKING OF COLLAGEN AFTER GLYCOSYLATION INVITRO
EVIDENCE FOR GLUCOSE-MEDIATED COVALENT CROSS-LINKING OF COLLAGEN AFTER GLYCOSYLATION INVITRO
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DOI：
10.1042/bj2250745
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发表时间：
1985-01-01
影响因子：
4.1
通讯作者：
BAILEY, AJ
中科院分区：
文献类型：
--
作者：
KENT, MJC;LIGHT, ND;BAILEY, AJ
兔前肢肌腱在35 ℃下孵育15或21天。C在8或24毫克/毫升的葡萄糖的存在下，显示出改变其化学，生物化学和机械特性。用葡萄糖处理的肌腱含有多达3倍的己糖基-赖氨酸和己糖基羟赖氨酸，如通过NaB 3 H4还原样品的测定所判断的那样。热收缩产生的力的测量结果显示，与对照组相比，糖基化肌腱显着增加，表明新的共价稳定键的形成。这一结论得到了完整肌腱和体外糖基化重建纤维溶解度降低以及溴化氰消化葡萄糖孵育肌腱肽图证据的支持。后者，当与对照肌腱的肽图谱相比，揭示了额外的高分子量肽材料的存在。这些肽似乎是通过一种新型的共价键交联的，这种共价键对温和的热和化学处理是稳定的。该系统在体外提供了一个易于控制的模型，用于研究糖尿病中非酶糖基化引起的胶原蛋白化学变化。
Rabbit forelimb tendons incubated for 15 or 21 days at 35.degree. C in the presence of 8 or 24 mg of glucose/ml were shown to change their chemical, biochemical and mechanical characteristics. The tendons treated with glucose contained up to 3 times as much hexosyl-lysine and hexosylhydroxylysine as did control tendons as judged by assay of NaB3H4-reduced samples. Measurement of the force generated on thermal contraction showed significant increases in glycosylated tendons compared with controls, indicating the formation of new covalent stabilizing bonds. This conclusion was supported by the decreased solubility of intact tendons and re-formed fibers glycosylated in vitro, and by the evidence from peptide maps of CNBr-digested glucose-incubated tendons. The latter, when compared with peptide maps of control tendons, revealed the presence of additional high-MW peptide material. These peptides appear to be cross-linked by a new type of covalent bond stable to mild thermal and chemical treatment. This system in vitro provides a readily controlled model for the study of the chemistry of changes brought about in collagen by non-enzymic glycosylation in diabetes.