反相高效液相色谱对CaBP_(21)的肽谱分析

PEPTIDE MAPPING OF CYANOGEN BROMIDE DIGESTS OF CaBP_21 BY REVERSEPHASE HIGH PERFQR MANGE LIQUID CHROMATOGRAPHY

对CaBP_(21),一种新发现的兔阑尾B淋巴细胞的钙结合蛋白,作手工Edman化学降解,结果表明其N末端封闭。取15nmole CaBP_(21)用溴化氰裂解,产物不经衍生,直接以RP-HPLC作肽谱分析。肽谱时间2h。得到两个BrCN肽解片段,其中C端片段可用作Edman顺序测定。同时证明了CaBP_(21)为相同亚基2聚体。

Manual Edman degradation of CaBP21, a newly discovered calcium-binding protein from rabbit appendix B lymphocytes, showed that its NH2 terminus is blocked. Peptide mapping of underivatized cyanogen bromide digests of CaBP21 was accomplished by reverse-phase high-performance liquid chromatog-raPhy (RP-HPLC) . High-precision analysis of 15 nanomolar quantities of peptides was achieved in 2 h ( recycle time ) . Two peptide fragments were obtained, with the C-terminal fragment fit for sequence analysis by Edman degradation. And it was also proved that CaBP21 consists of two identical subunits.