Expression and characterization of rat protein phosphatases-1 alpha, -1 gamma 1, -1 gamma 2, and -1 delta.
Four distinct cDNAs for rat protein phosphatase-1 have been isolated from rat tissues (Sasaki et al., Jpn. J. Cancer Res. 81, 1272-1280, 1990). These cDNAs encode proteins of highly similar sequence, the major differences being located at their N and C termini. In order to demonstrate that these cDNAs encode functional proteins and to investigate their enzymatic properties, it would be desirable to obtain purified preparations of these proteins. Using a system that was developed for the expression of rabbit muscle protein phosphatase-1 (Zhang et al., J. Biol. Chem. 267, 1484-1490, 1992) we have expressed these isoforms in Escherichia coli. The four recombinant isoforms were purified to near homogeneity and their properties were examined in terms of substrate specificity and sensitivity to okadaic acid and inhibitor-2.
Zhang
Z
Z
Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, Florida 33101.
Bai
G
G
Shima
M
M
Zhao
S
S
Nagao
M
M
Lee
E Y
EY
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000302:isolation & purification*; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D004988:Ethers, Cyclic / Q000494:pharmacology; D015870:Gene Expression; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D054645:Protein Phosphatase 1; D011506:Proteins / Q000494:pharmacology; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D014169:Transformation, Bacterial; D000097847:Protein Phosphatase Inhibitory Proteins
D004247:DNA; D004988:Ethers, Cyclic; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1; D011506:Proteins; D011994:Recombinant Proteins; C024833:protein phosphatase inhibitor-2; D000097847:Protein Phosphatase Inhibitory Proteins