Characterization of a high‐affinity Ins‐P4 (inositol 1,3,4,5‐tetrakisphosphate) receptor from brain by an anti‐peptide antiserum

FEBS Letters

Reiser, G.¹  Kalbacher, H.³  Lottspeich, F.²  Stricker, R.¹ 

[1] Institut für Neurobiochemie der Otto-von-Guericke Universität Magdeburg, Leipziger Str. 44, 39120 Magdeburg, Germany;Max-Planck-Institut für Biochemie, Martinsried, Germany;Physiologisch-Chemisches Institut der Eberhardt-Karls Universität, Tübingen, Germany

关键词: Inositolphosphate;    Signal transduction;    Affinity purification;    Ins-P4;    Ins(1;    3;    4;    5)P4;    d-myo inositol 1;    3;    4;    5-tetrakisphosphate;    KLH;    keyhole limpet hemocyanine;    PVDF;    polyvinylidene difluoride;   

DOI  :  10.1016/0014-5793(95)00822-Q

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown