Biosynthesis of collagen crosslinks: in vivo labelling of neonatal skin, tendon, and bone in rats.
Biosynthesis of collagen crosslinks: in vivo labelling of neonatal skin, tendon, and bone in rats.
复制标题
胶原交联的生物合成：大鼠新生皮肤、肌腱和骨骼的体内标记。
作者：
K. Reiser;J. Last
通过单次腹腔注射200 microCi [14 C]赖氨酸，在体内标记新生大鼠中的胶原交联。注射后30分钟至10周处死大鼠。在还原和水解后，通过HPLC分析整个皮肤、肌腱和骨的胶原交联含量。通过掺入[14 C]赖氨酸的放射性以及通过邻苯二甲醛柱后衍生化进行荧光测定，可视化交联和氨基酸。来自标记赖氨酸的14 C掺入到主要的双官能可还原交联中，即N6.6 '-异丙基-5，5'-二羟基赖氨酸正亮氨酸和N6.6 '-异丙基-5-羟基赖氨酸正亮氨酸，在注射后4 - 12小时之间增加最快，结果与其他人在体外研究交联生物合成时观察到的结果相似。将14 C掺入到四官能交联组氨酰羟基merodesmosine中进行得比双官能交联慢。对于研究的所有三种组织，在注射后3 - 5天，掺入各种交联的放射性量值达到表观恒定值。这些值在实验期间保持大致恒定，但肌腱中的HHMD除外，其显示在注射后8周和10周掺入的放射性增加。还通过测定邻苯二甲醛加合物的荧光对这些相同的交联进行了直接化学定量。我们的结论是，在体内标记的胶原交联可以进行研究，至少在快速生长的新生儿，单次注射放射性赖氨酸后。这些研究的结果支持以前的建议，由其他人的双功能交联的形成率的基础上，使用体外系统的研究。我们的研究结果进一步表明，形成的四功能可还原的交联histidinohydroxymerodesmosine在体内进行相对较快。最后，我们得出结论，这种标记的交联生物合成后显然是相当稳定的，这表明在大鼠的一生中的可观分数的胶原交联的代谢命运的研究的可能性。
Collagen crosslinks in neonatal rats were labelled in vivo by a single intraperitoneal injection of 200 microCi of [14C]lysine. Rats were killed at times ranging from 30 minutes to 10 weeks after injection. Whole skin, tendon, and bone were analyzed, after reduction and hydrolysis, for collagen crosslink content by HPLC. Crosslinks and amino acids were visualized by their incorporation of radioactivity from [14C]lysine and also fluorometrically by post-column derivatization with o-phthalaldehyde. The incorporation of 14C from labelled lysine into the principal difunctional reducible crosslinks, N6.6'-dehydro-5,5'-dihydroxylysinonorleucine and N6.6'-dehydro-5-hydroxylysinonorleucine, increased most rapidly between 4 and 12 hours after injection, results similar to those observed by others studying crosslink biosynthesis in vitro. Incorporation of 14C into the tetrafunctional crosslink histidinohydroxymerodesmosine proceeded more slowly than it did for the difunctional crosslinks. Values for the amount of radioactivity incorporated into the various crosslinks reached an apparent constant value between 3 and 5 days after injection for all three tissues studied. These values remained approximately constant for the duration of the experiment except for HHMD in tendon, which showed an increase in incorporated radioactivity at 8 and 10 weeks after injection. Direct chemical quantification of these same crosslinks by determination of the fluorescence of their o-phthalaldehyde adducts was also performed. We conclude that in vivo labelling of collagen crosslinks can be studied, at least in rapidly growing neonates, after a single injection of radioactive lysine. The results of such studies support previous suggestions by others about the rate of formation of difunctional crosslinks based upon studies using in vitro systems. Our results further suggest that formation of the tetrafunctional reducible crosslink histidinohydroxymerodesmosine proceeds relatively rapidly in vivo. Finally, we conclude that such labelled crosslinks are apparently quite stable after biosynthesis, suggesting the possibility of studies of the metabolic fate of collagen crosslinks over appreciable fractions of the lifetime of a rat.
骨的非矿化和矿化区室：吡啶啉在非矿化胶原蛋白中的作用。
DOI：
10.1016/0006-291x(83)91094-x
发表时间：
1983
影响因子：
3.1
作者：
Banes,AJ;Yamauchi,M;Mechanic,GL
通讯作者：
Mechanic,GL