Site-directed mutagenesis of prostatic acid phosphatase. Catalytically important aspartic acid 258, substrate specificity, and oligomerization

• Published in: The Journal of biological chemistry Vol. 269; no. 36; pp. 22642 - 22646
• Main Authors: Porvari, K S, Herrala, A M, Kurkela, R M, Taavitsainen, P A, Lindqvist, Y, Schneider, G, Vihko, P T
• Format: Journal Article
• Language: English
• Published: United States American Society for Biochemistry and Molecular Biology 09.09.1994
• Subjects: Acid Phosphatase - chemistry; Acid Phosphatase - isolation & purification; Acid Phosphatase - metabolism; Amino Acid Sequence; Animals; Aspartic Acid; Baculoviridae; Base Sequence; DNA Primers; Electrophoresis, Polyacrylamide Gel; Hydrogen-Ion Concentration; Immunoblotting; Kinetics; Macromolecular Substances; Male; Molecular Sequence Data; Moths; Mutagenesis, Site-Directed; Point Mutation; Polymerase Chain Reaction; Prostate - enzymology; Rats; Recombinant Proteins - chemistry; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Substrate Specificity; Transfection

At the active site of rat prostatic acid phosphatase (rPAP), residue Asp258 is a suitable candidate to act as an acid/base catalyst during phosphoester hydrolysis. It was changed to Asn, Ser, and Ala by site-directed mutagenesis. All these mutants were inactive, indicating that Asp258 may act as a p...