Purification and characterization of a guanosine diphosphatase activity from calf liver microsomal salt wash proteins.
A potent guanosine diphosphatase activity that hydrolyzes GDP to 5'-GMP + Pi has been isolated and purified from the salt wash proteins of calf liver microsomes. The purified enzyme, a monomeric protein of approximate Mr 46,000, possesses nucleotide substrate specificity since, among the nucleoside diphosphates and triphosphates tested, only GDP and UDP are hydrolyzed by the enzyme. The relative affinity of the enzyme for GDP is, however, much higher than for UDP. The effect of the enzyme on the binary complex formed between eukaryotic initiation factor 2 (eIF-2) and GDP has also been investigated. The enzyme neither hydrolyzes GDP bound to eIF-2 nor catalyzes the exchange of eIF-2-bound GDP with GTP even in the presence of Met-tRNAf. The enzyme, therefore, is presumably not involved in recycling of eIF-2 in eukaryotic polypeptide chain initiation reaction. The possible biological function of the enzyme in maintaining the cellular pool of GTP-GDP is discussed.
Raychaudhuri
P
P
Ghosh
S
S
Maitra
U
U
D000818:Animals; D002417:Cattle; D015852:Eukaryotic Initiation Factor-2; D006153:Guanosine Diphosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D008722:Methods; D008862:Microsomes, Liver / Q000201:enzymology*; D008970:Molecular Weight; D010448:Peptide Initiation Factors / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification*; D011506:Proteins / Q000378:metabolism; D011755:Pyrophosphatases; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D012358:RNA, Transfer, Met; D012269:Ribosomal Proteins / Q000032:analysis; D012965:Sodium Chloride
D015852:Eukaryotic Initiation Factor-2; D010448:Peptide Initiation Factors; D011506:Proteins; D012346:RNA, Transfer, Amino Acyl; D012358:RNA, Transfer, Met; D012269:Ribosomal Proteins; C024988:tRNA, formylmethionine-; D006153:Guanosine Diphosphate; D012965:Sodium Chloride; D006160:Guanosine Triphosphate; D010744:Phosphoric Monoester Hydrolases; D011755:Pyrophosphatases; C010357:guanosine-diphosphatase