Analysis of RNase A refolding intermediates by electrospray/mass spectrometry

FEBS Letters

Gennaro, Marino¹  Margherita, Ruoppolo¹  Claudia, Torella¹  Piero, Pucci¹ 

[1]Servizio Spettrometria di Massa, Via Pansini, 5, 80131 Naples, Italy

关键词: Electrospray/mass spectrometry;    Bovine pancreatic RNase A;    Refolding intermediate;    Alkylation;    DTNB;    5;    5′-dithio-bis(2-nitrobenzoic acid);    DTT;    reduced dithiothreitol;    EDTA;    ethylendiamine tetraacetic acid;    disodium salt;    GSSG;    oxidasedglutathione;    GSH;    reduced glutathione;    IAM;    iodoacetamine;    IAA;    iodoacetic acid;    Tris;    2-amino-2-(hydroxymethyl)-1;    3-propandiol;    ES/MS;    electrospray mass spectrometry;   

DOI  :  10.1016/0014-5793(94)00966-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Electrospray/mass spectrometry (ESIMS) was extensively used to obtain information on disulphide-containing intermediates formed during refolding of bovine pancreatic ribonuclease A. The analysis showed the existence of an equilibrated population of disulphide bonded intermediates, and indicates that intermediates containing two intramolecular S–S are predominant until late stages of the refolding process. Mixed disulphides with exogenous glutathione were also detected, supporting previous evidence of conformational restrictions on the ability of RNase A to form intramolecular disulphides. The results indicate that ES/MS is a suitable technique to detect and characterize refolding intermediates.

【 授权许可】

Unknown