Characterization of a high-affinity Ins-P4 (inositol 1,3,4,5-tetrakisphosphate) receptor from brain by an anti-peptide antiserum.
From a high-affinity Ins-P4 (inositol 1,3,4,5-P4) receptor purified from pig cerebellum, digested with the protease Lys C peptide sequences were obtained. Synthetic peptide-3 (19 amino acid residues) was used to generate an antiserum. Reaction of the affinity-purified antibodies with the purified pig receptor protein in ELISA or Western blot was completely inhibited by peptide-3. In cerebellar membranes, the antibodies clearly recognized the 42 kDa Ins-P4 receptor protein and two additional proteins (25 kDa, 37 kDa) which still have to be identified. The anti-peptide antibodies could selectively immunoprecipitate the Ins-P4 receptor protein. The antiserum was used (i) to demonstrate that in brain from different species (human, pig, beef, rat, mouse and sheep) a similar 42 kDa Ins-P4 receptor protein is contained, and (ii) to obtain indications for the existence of a related soluble form of the 42 kDa Ins-P4 receptor besides the membrane-associated receptor.
Stricker
R
R
Institut für Neurobiochemie der Otto-von-Guericke Universität Magdeburg, Germany.
Kalbacher
H
H
Lottspeich
F
F
Reiser
G
G
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000915:Antibody Affinity; D000918:Antibody Specificity; D015153:Blotting, Western; D001923:Brain Chemistry; D002417:Cattle; D002531:Cerebellum / Q000737:chemistry; D005591:Chemical Fractionation; D002845:Chromatography / Q000379:methods; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007106:Immune Sera / Q000494:pharmacology*; D007136:Immunoglobulins / Q000097:blood / Q000302:isolation & purification; D007295:Inositol Phosphates / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D011233:Precipitin Tests; D011817:Rabbits; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D013552:Swine
D000906:Antibodies; D007106:Immune Sera; D007136:Immunoglobulins; D007295:Inositol Phosphates; D010455:Peptides; D018160:Receptors, Cytoplasmic and Nuclear; C061107:inositol-1,3,4,5-tetrakisphosphate receptor; C081655:inositol-1,3,4,5-tetrakisphosphate