Nakagawa S, Tamakashi Y, Ishibashi Y, Kawase M, Taketomi S, Nishimura O, Fukuda T
Pharmaceutical Research Laboratories II, Takeda Chemical Industries, Ltd., Osaka, Japan.
Biochem Biophys Res Commun. 1994 May 16;200(3):1735-41. doi: 10.1006/bbrc.1994.1653.
The production of hPTH(1-34) by site specific chemical cleavage of [Cys35]hPTH(1-84) obtained by a biotechnology technique is described. The amino-peptide bond of S-cyanylated cysteine residues in peptides or proteins is specifically cleaved by exposure to an alkaline solution (Stark, G. R. (1977) Methods Enzymol, 47, 129-132). However, when applying this method to [Cys35]hPTH(1-84), we observed many by-products. Formation of these by-products was suppressed using a short reaction in 0.03N NaOH containing 6M urea at low temperature, and hPTH(1-34) was specifically produced. The product was indistinguishable from standard hPTH(1-34) with respect to chemical and biological characterization.
本文描述了通过生物技术获得的[Cys35]hPTH(1-84)的位点特异性化学裂解来生产hPTH(1-34)。肽或蛋白质中S-氰化半胱氨酸残基的氨基肽键通过暴露于碱性溶液而被特异性裂解（Stark, G. R. (1977) Methods Enzymol, 47, 129-132）。然而，当将此方法应用于[Cys35]hPTH(1-84)时，我们观察到许多副产物。通过在低温下于含6M尿素的0.03N NaOH中进行短时间反应抑制了这些副产物的形成，并特异性地产生了hPTH(1-3)。就化学和生物学特性而言，该产物与标准hPTH(1-34)无法区分。
