Receptor for myo-inositol trisphosphate from the microsomal fraction of Vigna radiata.
The microsomal fraction from mung-bean (Vigna radiata) hypocotyl was found to contain Ins (1,4,5)P3- and Ins(2,4,5)P3-binding activity. Preincubation of the microsomal fraction with thiol-containing reagents reduced specific InsP3 binding. A single class of binding site with a Kd value of 1.5 nM and Bmax. of 1.1 pmol/mg of protein was detected. Other myo-inositol phosphates exhibited little affinity for this protein. The binding protein was purified to homogeneity and the molecular mass of the native form recorded as 400 kDa. However, under denaturing conditions the molecular mass was 110 kDa, suggesting that the protein is a homotetramer. That this protein is associated with Ca2+ release was confirmed by including it in proteoliposomes and adding Ins(1,4,5)P3 or Ins(2,4,5)P3. The affinity of Ins(1,4,5)P3 is 3-fold higher than that of Ins(2,4,5)P3. The binding affinity of InsP3 is also reflected in the extent of Ca2+ released from the microsomal fraction. Heparin inhibits binding of InsP3 to the protein, the K1/2 being 0.26 microM. It is also shown that the protein acts as a receptor for InsP3 with characteristics of high affinity and low density.
Biswas
S
S
Department of Biochemistry, Bose Institute, Calcutta, India.
Dalal
B
B
Sen
M
M
Biswas
B B
BB
D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000302:isolation & purification* / Q000378:metabolism; D006493:Heparin / Q000494:pharmacology; D053496:Inositol 1,4,5-Trisphosphate Receptors; D008861:Microsomes / Q000378:metabolism; D008970:Molecular Weight; D010944:Plants / Q000378:metabolism*; D011869:Radioligand Assay; D018160:Receptors, Cytoplasmic and Nuclear / Q000302:isolation & purification* / Q000378:metabolism
D015220:Calcium Channels; D053496:Inositol 1,4,5-Trisphosphate Receptors; D018160:Receptors, Cytoplasmic and Nuclear; D006493:Heparin; D002118:Calcium