Conformational stability and activity of ribonuclease T1 and mutants: Gln super(25) arrow right Lys, Glu super(53) arrow right Ala, and the double mutant

Ribonuclease T1 (RNase T1) and mutants Gln super(25) arrow right Lys, Glu super(53) arrow right Ala, and the double mutant were prepared from a chemically synthesized gene, cloned and expressed in Escherichia coli . The wild-type RNase T1 prepared from the cloned gene was identical in every function...

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Bibliographic Details
Published in:The Journal of biological chemistry 1989-01, Vol.264 (20), p.11621-11625
Main Authors: Shirley, BA, Stanssens, P, Steyaert, J, Pace, C N
Format: Article
Language:English
Subjects:
Aspergillus oryzae
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Summary:Ribonuclease T1 (RNase T1) and mutants Gln super(25) arrow right Lys, Glu super(53) arrow right Ala, and the double mutant were prepared from a chemically synthesized gene, cloned and expressed in Escherichia coli . The wild-type RNase T1 prepared from the cloned gene was identical in every functional and physical property examined to RNase T1 prepared from Aspergillus oryzae . Urea and thermal unfolding experiments show that Gln super(25) arrow right Lys is 0.9 kcal/mol more stable and Glu super(58) arrow right Ala is 0.8 kcal/mol less stable than wild-type RNase T1. In the double mutant, these contributions cancel and the stability does not differ significantly from that of wild-type RNase T1.
ISSN:0021-9258