The disulphide bond pattern of bitistatin, a disintegrin isolated from the venom of the viper Bitis arietans

FEBS Letters

Calvete, Juan J⁵  Raida, Manfred³  Niewiarowski, Stefan²  Romero, Antonio⁴  Schrader, Michael³  McLane, Mary Ann¹ 

[1] Department of Medical Technology, University of Delaware, Newark, DE, USA;Temple University School of Medicine, Philadelphia, PA, USA;Niedersächsiches Institut für Peptid-Forschung (IPF) GmbH, Hannover, Germany;Centro de Investigaciones Biológicas, CSIC, Madrid, Spain;Institut für Reproduktionsmedizin, Tierärztliche Hochschule, Bünteweg 15, D-30559 Hannover-Kirchrode, Germany

关键词: Disulfide bond;    Disintegrin;    Bitistatin;    RGD peptide;    Snake venom;    Mass spectrometry;    Oxalic acid degradation;   

DOI  :  10.1016/S0014-5793(97)01203-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The disulphide bond pattern of the long disintegrin bitistatin (83 amino acids, 14 cysteines) was established using structural information gathered by amino acid analysis, N-terminal sequencing, and molecular mass determination of fragments isolated by reversed-phase HPLC after polypeptide degradation with trypsin and oxalic acid. A computer program was used to calculate all possible combinations of disulphide-bonded peptides matching the mass spectrometric data, and the output was filtered using compositional and sequence data. Disulphide bonds between cysteines 16–34, 18–29, 28–51, 42–48, 47–72, and 60–79 are conserved in medium-long disintegrins flavoridin and kistrin (70 amino acids, 12 cysteines), and the two cysteine residues at positions 5 and 24 found in bitistatin but not in other disintegrin molecules are disulphide-bridged. This linkage creates an extra, large loop, which, depending on whether the NMR structure of flavoridin or kistrin is used for modelling the structure of bitistatin, lies opposite or nearly parallel, respectively, to the biologically active RGD-containing loop.

【 授权许可】

Unknown