Molecular cloning and expression of a human brain inositol 1,4,5-trisphosphate 3-kinase.
A human hippocampus cDNA library was screened by hybridization with a rat brain inositol 1,4,5-trisphosphate (InsP3) 3-kinase cDNA. Sequencing of three overlapping clones identified a 1383 bp open reading frame encoding a 461 amino acid protein with a calculated molecular weight of 50988. The coding amino acid sequence showed an overall 93% similarity with the sequence of rat brain InsP3 3-kinase. The cDNA insert of one isolated partial clone (i.e. hh 26) was in frame with the beta galactosidase fragment fused to it as a Bluescript plasmid; it displayed InsP3 3-kinase activity when expressed in Escherichia coli (E. coli). Biochemical characterization of human brain InsP3 3-kinase by SDS polyacrylamide gel electrophoresis and regeneration of enzyme activity reveals three active fractions with apparent Mr of 58,000-64,000, 45,000-50,000 and 37,000-39,000.
Takazawa
K
K
Institut de Recherche Interdisciplinaire (IRIBHN), Université Libre de Bruxelles, Belgium.
Perret
J
J
Dumont
J E
JE
Erneux
C
C
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D015723:Gene Library; D006624:Hippocampus / Q000201:enzymology*; D006801:Humans; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010770:Phosphotransferases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor); D051381:Rats; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
D011994:Recombinant Proteins; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C051038:Inositol 1,4,5-trisphosphate 3-kinase