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Analysis of RNase A refolding intermediates by electrospray/mass spectrometry
Author(s) -
Claudia Torella,
Margherita Ruoppolo,
Gennaro Marino,
Piero Pucci
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)00966-x
Subject(s) - chemistry , bovine pancreatic ribonuclease , intramolecular force , mass spectrometry , rnase p , electrospray mass spectrometry , ribonuclease , glutathione , electrospray , biochemistry , stereochemistry , chromatography , enzyme , rna , gene
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.