Covalent protein crosslinks: general detection, quantitation, and characterization via modification with diphenylborinic acid.
Covalent protein crosslinks: general detection, quantitation, and characterization via modification with diphenylborinic acid.
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共价蛋白质交联：通过二苯基硼酸修饰进行一般检测、定量和表征。
DOI：
10.1006/abio.1994.1122
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发表时间：
1994
影响因子：
2.9
通讯作者：
Gallop,PM
中科院分区：
文献类型：
--
作者：
Graham,L;Gallop,PM
蛋白质的渐进性交联似乎是衰老细胞和组织中的一种普遍现象。交联蛋白可以形成不溶的聚集体，随着更多的交联物的形成，这种聚集体对蛋白质降解的抵抗力越来越强。然而，随着年龄的增长，大多数证据是间接的，对其中涉及的化学机制知之甚少。因此，我们开发了一种从蛋白质水解物中检测和分离任何类型的稳定的共价交联物的方法，该方法不需要事先知道任何可能存在的交联物的分子结构(S)。它利用了二苯基硼酸试剂对α-氨基酸基团的专一性以及交联物的层析性质和紫外吸收。该方法使用了8种不同的胶原和纤维蛋白交联物，并给出了检测蛋白质水解物中任何类型交联物的一般程序。
Progressive crosslinking of proteins appears to be a general phenomenon in aging cells and tissues. Crosslinked proteins can form insoluble aggregates which become increasingly resistant to proteolysis as more crosslinks form. However, most evidence for progressive crosslinking with age is indirect, and little is known about the chemical mechanisms involved. We have therefore developed a method for detection and isolation of any type of stable covalent crosslink from protein hydrolysates which requires no prior knowledge of the molecular structure of whatever crosslink(s) may be present. It utilizes the specificity of the diphenylborinic acid reagent for α-amino acid groups and the chromatographic properties and uv absorbance of the crosslink derivatives. The method is demonstrated using eight different crosslinks from collagen and fibrin, and a general procedure is given for detection of any type of crosslink in a protein hydrolysate.