Anti‐head and anti‐tail antibodies against distinct epitopes in the catalytic subunit of protein kinase A Use in the study of the kinase splitting membranal proteinase KSMP

FEBS Letters

Chestukhin, Anton¹  Litovchick, Larisa¹  Batkin, Misha¹  Shaltiel, Shmuel¹ 

[1] Department of Biological Regulation, The Weizmann Institute of Science, Rehovot 76100, Israel

关键词: Anti-peptide antibodies;    Inhibitory antibodies;    KSMP;    Protein kinase A;    Tails of protein kinases;    C;    catalytic subunit of cAMP-dependent protein kinase;    DAB;    3;    3′-diaminobenzidine;    KLH;    keyhole limpet hemocyanin;    KSMP;    kinase-splitting membranal proteinase;    C′;    KSMP-cleaved C;    PKA;    protein kinase A (cAMP-dependent protein kinase);   

DOI  :  10.1016/0014-5793(96)00178-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Protein kinases share a considerable sequence homology in their catalytic core (residues 40–300 in PKA). Each core is flanked by “head” and “tail” segments at its amino- and carboxy-termini, which are different in the various kinases. These end segments may play an important role in creating the preferential affinity of each kinase for its physiological substrates or regulatory ligands. Here we describe three anti-peptide antibodies (αP-1, αP-2, and αP-3) that specifically recognize the head and tail segments of the catalytic subunit (C) of PKA. (i) αP-1 (against ⁶A-K²³) react with C when denatured but not when in its native structure; (ii) αP-2 (against ³¹⁹K-I³³⁵), bind to the site in C cleaved by the kinase splitting membranal proteinase (KSMP) and inhibit this cleavage of C; (iii) αP-3 (against ³³⁸S-F³⁵⁰) react with C but not with the KSMP cleavage product C′, useful for detecting a KSMP-like activity in different tissues and subcellular loci. The combined use of the antibodies described here provides a strict definition of C, and thus a high degree of fidelity in its biorecognition.

【 授权许可】

Unknown