D-myo-inositol 1,4,5-trisphosphate binding domain of phospholipase C-delta 1.
D-myo-Inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) binding domain of phospholipase C-delta 1 (PLC-delta 1) was determined by examining binding activity of the synthetic peptide corresponding to residues 30-43 of PLC-delta 1. The peptide coupled with carrier proteins such as keyhole limpet hemocyanin or bovine serum albumin bound Ins(1,4,5)P3, whereas a scrambled peptide with the same amino acids did not do so. Polyclonal antibody against the peptide was examined to determine whether it would cause inhibition of the Ins(1,4,5)P3 binding to PLC-delta 1. Fab fragment of antibody to the peptide did inhibit binding to PLC-delta 1, in a dose-dependent manner. Thus it seems likely that the region of residues 30-43 of PLC-delta 1 is responsible for the binding of Ins(1,4,5)P3.
Hirata
M
M
Department of Biochemistry, Faculty of Dentistry, Kyushu University, Fukuoka, Japan.
Kanematsu
T
T
Sakuma
K
K
Koga
T
T
Watanabe
Y
Y
Ozaki
S
S
Yagisawa
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001665:Binding Sites; D002352:Carrier Proteins / Q000378:metabolism; D002417:Cattle; D006433:Hemocyanins / Q000378:metabolism; D066298:In Vitro Techniques; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000235:genetics / Q000378:metabolism; D054803:Phospholipase C delta; D051381:Rats; D012710:Serum Albumin, Bovine / Q000378:metabolism; D010738:Type C Phospholipases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D000906:Antibodies; D002352:Carrier Proteins; D007527:Isoenzymes; D010446:Peptide Fragments; D012710:Serum Albumin, Bovine; D015544:Inositol 1,4,5-Trisphosphate; D006433:Hemocyanins; D010738:Type C Phospholipases; D054803:Phospholipase C delta; C032808:keyhole-limpet hemocyanin