The catalytic subunit of Dictyostelium cAMP-dependent protein kinase - Role of the N-terminal domain and of the C-terminal residues in catalytic activity and stability

被引:33
作者
Etchebehere, LC
VanBemmelen, MXP
Anjard, C
Traincard, F
Assemat, K
Reymond, C
Veron, M
机构
[1] INST PASTEUR,UNITE REGULAT ENZYMAT ACT CELLULAIRES,CNRS,UMR 321,F-75745 PARIS 15,FRANCE
[2] UNIV SAO PAULO,INST QUIM,DEPT BIOQUIM,BR-01498 SAO PAULO,BRAZIL
[3] INST PASTEUR,UNITE IMMUNOL STRUCT,F-75745 PARIS 15,FRANCE
[4] UNIV LAUSANNE,INST HISTOL,LAUSANNE,SWITZERLAND
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1997年 / 248卷 / 03期
关键词
cAMP-dependent protein kinase; cAMP; protein kinase A; Dictyostelium discoideum;
D O I
10.1111/j.1432-1033.1997.t01-2-00820.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The C subunit of Dictyostelium cAMP-dependent protein kinase (PKA) is unusually large (73 kDa) due to the presence of 330 amino acids N-terminal to the conserved catalytic core. the sequence following the core, including a C-terminal -Phe-Xaa-Xaa-Phe-COOH motif, is highly conserved. We have characterized the catalytic activity and stability of C subunits mutated in sequences outside the catalytic core and we have analyzed their ability to interact with the R subunit and with the heat-stable protein-kinase inhibitor PKI. Mutants carrying deletions in the N-terminal domain displayed little difference in their kinetic properties and retained their capacity to be inhibited by R subunit and by PKI. In contrast, the mutation of one or both of the phenylalanine residues in the C-terminal motif resulted in a decrease of catalytic activity and stability of the proteins. Inhibition by the R subunit or by PKI were however unaffected. Sequence-comparison analysis of other protein kinases revealed that a -Phe-Xaa-Xaa-Phe- motif is present in many Ser/Thr protein kinases, although its location at the very end of the polypeptide is a particular feature of the PKA family. We propose that the presence of this motif may serve to identify isoforms of protein kinases.
引用
收藏
页码:820 / 826
页数:7
相关论文
共 40 条
[1]   SEQUENCE AND EXPRESSION OF CHICKEN AND MOUSE RSK - HOMOLOGS OF XENOPUS-LAEVIS RIBOSOMAL S6 KINASE [J].
ALCORTA, DA ;
CREWS, CM ;
SWEET, LJ ;
BANKSTON, L ;
JONES, SW ;
ERIKSON, RL .
MOLECULAR AND CELLULAR BIOLOGY, 1989, 9 (09) :3850-3859
[2]   AN UNUSUAL CATALYTIC SUBUNIT FOR THE CAMP-DEPENDENT PROTEIN-KINASE OF DICTYOSTELIUM-DISCOIDEUM [J].
ANJARD, C ;
ETCHEBEHERE, L ;
PINAUD, S ;
VERON, M ;
REYMOND, CD .
BIOCHEMISTRY, 1993, 32 (37) :9532-9538
[3]   MOLECULAR-CLONING OF A TISSUE-SPECIFIC PROTEIN-KINASE (C-GAMMA) FROM HUMAN TESTIS - REPRESENTING A 3RD ISOFORM FOR THE CATALYTIC SUBUNIT OF CAMP-DEPENDENT PROTEIN-KINASE [J].
BEEBE, SJ ;
OYEN, O ;
SANDBERG, M ;
FROYSA, A ;
HANSSON, V ;
JAHNSEN, T .
MOLECULAR ENDOCRINOLOGY, 1990, 4 (03) :465-475
[4]   2 CATALYTIC SUBUNITS OF CAMP-DEPENDENT PROTEIN-KINASE GENERATED BY ALTERNATIVE RNA SPLICING ARE EXPRESSED IN APLYSIA NEURONS [J].
BEUSHAUSEN, S ;
BERGOLD, P ;
STURNER, S ;
ELSTE, A ;
ROYTENBERG, V ;
SCHWARTZ, JH ;
BAYLEY, H .
NEURON, 1988, 1 (09) :853-864
[5]  
BOGOSIAN G, 1990, RIBOSOME, P546
[6]   PHOSPHOTRANSFERASE AND SUBSTRATE BINDING MECHANISM OF THE CAMP-DEPENDENT PROTEIN-KINASE CATALYTIC SUBUNIT FROM PORCINE HEART AS DEDUCED FROM THE 2.0 ANGSTROM STRUCTURE OF THE COMPLEX WITH MN2+ ADENYLYL IMIDODIPHOSPHATE AND INHIBITOR PEPTIDE PKI(5-24) [J].
BOSSEMEYER, D ;
ENGH, RA ;
KINZEL, V ;
PONSTINGL, H ;
HUBER, R .
EMBO JOURNAL, 1993, 12 (03) :849-859
[7]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[8]  
BUECHLER YJ, 1993, J BIOL CHEM, V268, P16495
[9]   ISOLATION OF 2 GENES ENCODING PUTATIVE PROTEIN-KINASES REGULATED DURING DICTYOSTELIUM-DISCOIDEUM DEVELOPMENT [J].
BURKI, E ;
ANJARD, C ;
SCHOLDER, JC ;
REYMOND, CD .
GENE, 1991, 102 (01) :57-65
[10]   Functional malleability of the carboxyl-terminal tail in protein kinase A [J].
Chestukhin, A ;
Litovchick, L ;
Schourov, D ;
Cox, S ;
Taylor, SS ;
Shaltiel, S .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (17) :10175-10182