Williamson R A, Smith B J, Angal S, Murphy G, Freedman R B
Biological Laboratory, University of Kent, Canterbury, UK.
Biochim Biophys Acta. 1993 Jun 24;1164(1):8-16. doi: 10.1016/0167-4838(93)90105-z.
Tryptic digests of recombinant TIMP-1 have been resolved on reverse-phase HPLC and the major peaks identified by N-terminal sequencing. This procedure accounted for the entire molecule, except two short peptides of 2 and 4 amino acids in length. The peptide map was used to (i), characterize an insoluble 'core' peptide seen on digestion of TIMP-1 in non-reducing conditions; (ii), confirm the structure of delta 127-184TIMP-1, a recently described truncated form of the TIMP-1 molecule; (iii), identify exposed regions of the intact and truncated TIMP-1 molecules by measuring the rate of tryptic peptide release and (iv), locate sites of aberrant proteolysis seen when recombinant human TIMP-1 was purified at large scale.
重组组织金属蛋白酶抑制剂-1（TIMP-1）的胰蛋白酶消化产物已通过反相高效液相色谱法进行分离，并通过N端测序鉴定了主要峰。该方法解释了整个分子的情况，但有两个长度分别为2个和4个氨基酸的短肽除外。肽图被用于：（i）表征在非还原条件下TIMP-1消化时出现的不溶性“核心”肽；（ii）确认δ127-184TIMP-1（TIMP-1分子最近描述的一种截短形式）的结构；（iii）通过测量胰蛋白酶肽释放速率来鉴定完整和截短的TIMP-1分子的暴露区域；以及（iv）定位大规模纯化重组人TIMP-1时出现异常蛋白水解的位点。
