Serine/threonine protein phosphatases and a protein phosphatase 1 inhibitor from Neurospora crassa.
The major spontaneously active serine/threonine (Ser/Thr) protein phosphatase activities in N. crassa wild type (FGSC 424) were type-1 (PP1), type-2A (PP2A) and type-2C (PP2C). PP1 and PP2C predominantly dephosphorylated phosphorylase a and casein, respectively. PP2A acted on both substrates, but was two-fold more active against casein. PP1 activity was inhibited by protamine, heparin, okadaic acid (IC50 50 nM) and mammalian inhibitor-1 (IC50 2 nM). On the other hand. PP2A activity was inhibited by much lower concentrations of okadaic acid (IC50 0.2 nM) and also by protamine, but not by heparin or inhibitor-1. About 80% of total PP1 activity was associated with the particulate fraction and could be partially extracted with 0.5 M NaCl. Seventy and ninety percent of PP2A and PP2C activities, respectively, were found in the soluble fraction. In addition we have partially purified an acid and thermostable PP1 inhibitor which effectively inhibits both N. crassa and mammalian PP1.
Zapella
P D
PD
Departamento de Bioquímica, Universidade de São Paulo, Brasil.
da-Silva
A M
AM
da-Costa-Maia
J C
JC
Terenzi
H F
HF
D000818:Animals; D002845:Chromatography; D009492:Neurospora crassa / Q000201:enzymology*; D019319:Okadaic Acid / Q000494:pharmacology*; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D054645:Protein Phosphatase 1; D051381:Rats; D012694:Serine / Q000378:metabolism*; D013379:Substrate Specificity; D013912:Threonine / Q000378:metabolism*
D019319:Okadaic Acid; D013912:Threonine; D012694:Serine; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1