Cloning and expression of a human placenta inositol 1,3,4,5-tetrakisphosphate and phosphatidylinositol 3,4,5-trisphosphate 5-phosphatase.
Distinct inositol and phosphatidylinositol polyphosphate 5-phosphatases have recently been cloned. Primers were designated coding for highly conserved amino acid regions that are shared between sequences of 5-phosphatases. We used degenerate primers to amplify polymerase chain reaction products from rat brain cDNA. A product with a novel sequence was identified and used to clone a 4.9 kb cDNA from human placenta cDNA libraries (hp51CN). COS-7 cells transfected with a C-terminal truncated form of this cDNA showed an increase in Ins(1,3,4,5)P4 and PtdIns(3,4,5)P3 hydrolyzing activity, but not in Ins(1,4,5)P3 5-phosphatase. Enzymatic activity was inhibited in the presence of 2,3-bisphosphoglycerate and p-hydroxymercuribenzoate. The presence of an SH2 domain and proline-rich sequence motifs within hp51CN suggests that this 5-phosphatase interacts with various proteins in signal transduction.
Drayer
A L
AL
Interdisciplinary Research Institute (IRIBHN), Université Libre de Bruxelles, Belgium.
Pesesse
X
X
De Smedt
F
F
Woscholski
R
R
Parker
P
P
Erneux
C
C
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000201:enzymology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D017124:Conserved Sequence; D017931:DNA Primers; D005260:Female; D015870:Gene Expression; D006801:Humans; D000072181:Inositol Polyphosphate 5-Phosphatases; D007700:Kinetics; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010744:Phosphoric Monoester Hydrolases / Q000096:biosynthesis* / Q000378:metabolism; D010920:Placenta / Q000201:enzymology*; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D013379:Substrate Specificity; D014162:Transfection; D018909:src Homology Domains
D017931:DNA Primers; D011994:Recombinant Proteins; C096024:phosphatidylinositol-3,4,5-trisphosphate 5-phosphatase; D010744:Phosphoric Monoester Hydrolases; D000072181:Inositol Polyphosphate 5-Phosphatases