Refolding of RNAse A at high concentrations: identification of non-native species.
In this paper, we present an analysis of the soluble species formed on refolding of RNase A at various concentrations, in order to characterize these species with respect to structure and activities. Studies were carried out using reverse-phase high-performance liquid chromatography, circular dichroism, chromatography and ultracentrifugation. At all concentrations of protein used, RNase A refolded to the native form, together with formation of non-native species. These non-native species are either misfolded monomers or aggregates; the percentage of such species increases with increasing concentration of enzyme. Such aggregation appears to be a non-random process governed by intermolecular disulfide crosslinking between monomers. These results reaffirm the principle that the information for folding of the protein is encoded in the amino acid sequence itself.
Kumar
T K
TK
Centre for Cellular and Molecular Biology, Hyderabad, India.
Gopalakrishna
K
K
Ramakrishna
T
T
Pandit
M W
MW
D002851:Chromatography, High Pressure Liquid; D017510:Protein Folding; D012259:Ribonuclease, Pancreatic / Q000032:analysis / Q000737:chemistry*
D012259:Ribonuclease, Pancreatic