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Totally synthetic crystalline ribonuclease A
Author(s) -
Yajima Haruaki,
Fujii Nobutaka
Publication year - 1981
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.1981.360200910
Subject(s) - chemistry , bovine pancreatic ribonuclease , s tag , thioanisole , rnase p , ribonuclease , enzyme , chromatography , ion chromatography , biochemistry , combinatorial chemistry , organic chemistry , catalysis , rna , gene
Improved chemical synthesis of bovine pancreatic ribonuclease (RNase) A was achieved by applying a new deprotecting procedure with trifluoromethanesulfonic acid–thioanisole in combination with a modified air‐oxidation procedure with glutathione for the disulfide formation. After purifications by affinity chromatography, followed by ion‐exchange chromatography, a protein with the full enzymatic activity was obtained and subsequently crystallized from aqueous ethanol according to Kunitz. A totally synthetic enzyme with full RNase A activity was thus obtained in a crystalline form for the first time.