Han Y N, Kato H, Iwanaga S, Suzuki T
J Biochem. 1976 Jun;79(6):1201-22. doi: 10.1093/oxfordjournals.jbchem.a131175.
On incubation of bovine plasma high-molecular-weight (HMW) kininogen with purified plasma kallikrein [EC 3.4.21.8], a large glycopeptide fragment and the vasoactive peptide, bradykinin, were initially liberated; the former, named fragment 1-2, was subsequently cleaved into fragment 1 (glycopeptide) and the previously established fragment 2 (histidine-rich peptide). The isolated fragment 1-2 contained a total of 108 to 110 amino acid residues and carbohydrates, and the amino-terminal sequence Ser-Val-Gln was established. The other fragment, fragment 1, consisted of a total of 69 amino acid residues with serine and arginine (and lysine) at the amino and carboxyl termini, respectively. It contained eleven residues each of histidine and glycine, together with an oligosaccharide chain consisting of galactosamine, hexose and sialic acid. The complete amino acid sequence of fragment 1 was determined by Edman degradation and standard enzymatic and chemical techniques. These results established the following sequence: H-Ser-Val-Gln-Val-Met-Lys-Thr-Glu-Gly-Ser-Thr-Pro/Thr-Val-Ser(CHO)-Val/Leu-Pro-His-Ser-Ala-Met-Ser-Pro-Val-Gln-Asp-Glu-Glu-Arg-Asp-Ser-Gly-Lys-Glu-Gln-Gly-Pro-Thr-His-Gly-His-Gly-Trp-Asp-His-Gly-Lys-Gln-Ile-Lys-Leu-His-Gly-Leu-Gly-Leu-Gly-His-Lys-His-Lys-His-Asp-Gln-Gly-His-Gly-His-His-Lys/ArgOH.
将牛血浆高分子量(HMW)激肽原与纯化的血浆激肽释放酶[EC 3.4.21.8]一起温育时,最初会释放出一个大的糖肽片段和血管活性肽缓激肽;前者命名为片段1-2,随后被切割成片段1(糖肽)和先前确定的片段2(富含组氨酸的肽)。分离出的片段1-2总共包含108至110个氨基酸残基和碳水化合物,并确定了氨基末端序列Ser-Val-Gln。另一个片段,片段1,总共由69个氨基酸残基组成,氨基末端和羧基末端分别为丝氨酸和精氨酸(以及赖氨酸)。它含有11个组氨酸残基和11个甘氨酸残基,以及一条由半乳糖胺、己糖和唾液酸组成的寡糖链。通过埃德曼降解以及标准的酶促和化学技术确定了片段1的完整氨基酸序列。这些结果确定了以下序列:H-Ser-Val-Gln-Val-Met-Lys-Thr-Glu-Gly-Ser-Thr-Pro/Thr-Val-Ser(CHO)-Val/Leu-Pro-His-Ser-Ala-Met-Ser-Pro-Val-Gln-Asp-Glu-Glu-Arg-Asp-Ser-Gly-Lys-Glu-Gln-Gly-Pro-Thr-His-Gly-His-Gly-Trp-Asp-His-Gly-Lys-Gln-Ile-Lys-Leu-His-Gly-Leu-Gly-Leu-Gly-His-Lys-His-Lys-His-Asp-Gln-Gly-His-Gly-His-His-Lys/ArgOH。