Tang J, Sepulveda P, Marciniszyn J, Chen K C, Huang W Y, Tao N, Liu D, Lanier J P
Proc Natl Acad Sci U S A. 1973 Dec;70(12):3437-9. doi: 10.1073/pnas.70.12.3437.
As the culmination of several years of experiments, we propose a complete amino-acid sequence for porcine pepsin, an enzyme containing 327 amino-acid residues in a single polypeptide chain. In the sequence determination, the enzyme was treated with cyanogen bromide. Five resulting fragments were purified. The amino-acid sequence of four of the fragments accounted for 290 residues. Because the structure of a 37-residue carboxyl-terminal fragment was already known, it was not studied. The alignment of these fragments was determined from the sequence of methionyl-peptides we had previously reported. We also discovered the locations of activesite aspartyl residues, as well as the pairing of the three disulfide bridges. A minor component of commercial crystalline pepsin was found to contain two extra amino-acid residues, Ala-Leu-, at the amino-terminus of the molecule. This minor component was apparently derived from a different site of cleavage during the activation of porcine pepsinogen.
经过数年实验的最终成果,我们提出了猪胃蛋白酶完整的氨基酸序列,该酶在一条单多肽链中含有327个氨基酸残基。在序列测定过程中,该酶用溴化氰处理。得到的五个片段被纯化。其中四个片段的氨基酸序列占290个残基。由于一个37个残基的羧基末端片段的结构已经知晓,因此未对其进行研究。这些片段的比对是根据我们之前报道的甲硫氨酰肽的序列确定的。我们还发现了活性位点天冬氨酸残基的位置以及三个二硫键的配对情况。发现商业结晶胃蛋白酶的一个次要成分在分子的氨基末端含有两个额外的氨基酸残基,即丙氨酸-亮氨酸-。这个次要成分显然来自猪胃蛋白酶原激活过程中不同的切割位点。