RECONSTITUTION OF HETEROLOGOUS AND CHIMERIC CASEIN KINASE-II WITH RECOMBINANT SUBUNITS FROM HUMAN AND DROSOPHILA - IDENTIFICATION OF SPECIES-SPECIFIC DIFFERENCES IN THE BETA-SUBUNIT

被引:1
作者
LIN, WJ [1 ]
JAKOBI, R [1 ]
TRAUGH, JA [1 ]
机构
[1] UNIV CALIF RIVERSIDE, DEPT BIOCHEM, RIVERSIDE, CA 92521 USA
来源
JOURNAL OF PROTEIN CHEMISTRY | 1994年 / 13卷 / 02期
关键词
CASEIN KINASE; PROTEIN KINASE; RECOMBINANT PROTEIN; RECONSTITUTION; PROTEIN RENATURATION;
D O I
10.1007/BF01891979
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
Casein kinase IT is composed of two catalytic (alpha) and two regulatory (beta) subunits, the amino acid sequences of the alpha and beta subunits are highly conserved between species. To examine whether heterologous casein kinase II could be formed, recombinant alpha and beta subunits from human and Drosophila were reconstituted from inclusion bodies. Casein kinase II containing either human alpha and Dposophila beta or Drosophila alpha and human beta subunits exhibited enzymatic properties similar to those of the homologous holoenzymes with regard to specific activity, salt optima, and autophosphorylation. However, renaturation and reconstitution of casein kinase II was dependent on the type of beta subunits and the redox conditions, with the Drosophila beta subunits requiring more reduced conditions. Chimeric beta subunits prepared from human and Drosophila cDNA revealed that the N-terminal region was responsible for the requirement for the reduced redox state during renaturation. The N-terminal region also affected solubility and electrophoretic mobility of the beta subunit.
引用
收藏
页码:217 / 225
页数:9
相关论文
共 32 条
[1]  
BIDWAI AP, 1992, J BIOL CHEM, V267, P18790
[2]   EXPRESSION AND PURIFICATION OF THE ALPHA-SUBUNIT AND BETA-SUBUNIT OF DROSOPHILA CASEIN KINASE-II USING A BACULOVIRUS VECTOR [J].
BIRNBAUM, MJ ;
WU, JG ;
OREILLY, DR ;
RIVERAMARRERO, CA ;
HANNA, DE ;
MILLER, LK ;
GLOVER, CVC .
PROTEIN EXPRESSION AND PURIFICATION, 1992, 3 (02) :142-150
[3]   THE BETA-SUBUNIT OF CASEIN KINASE-II - CLONING OF CDNAS FROM MURINE AND PORCINE ORIGIN AND EXPRESSION OF THE PORCINE SEQUENCE AS A FUSION PROTEIN [J].
BOLDYREFF, B ;
PIONTEK, K ;
SCHMIDTSPANIOL, I ;
ISSINGER, OG .
BIOCHIMICA ET BIOPHYSICA ACTA, 1991, 1088 (03) :439-441
[4]   ISOLATION, SEQUENCING, AND DISRUPTION OF THE CKA1 GENE ENCODING THE ALPHA-SUBUNIT OF YEAST CASEIN KINASE-II [J].
CHENWU, JLP ;
PADMANABHA, R ;
GLOVER, CVC .
MOLECULAR AND CELLULAR BIOLOGY, 1988, 8 (11) :4981-4990
[5]  
COCHET C, 1983, J BIOL CHEM, V258, P1403
[6]  
FILHOL O, 1992, J BIOL CHEM, V267, P20577
[7]   POLYAMINE BINDING-ACTIVITY OF CASEIN KINASE-II [J].
FILHOL, O ;
COCHET, C ;
DELAGOUTTE, T ;
CHAMBAZ, EM .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1991, 180 (02) :945-952
[8]   ISOLATION AND CHARACTERIZATION OF RECOMBINANT HUMAN CASEIN KINASE-II SUBUNITS ALPHA AND BETA FROM BACTERIA [J].
GRANKOWSKI, N ;
BOLDYREFF, B ;
ISSINGER, OG .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1991, 198 (01) :25-30
[9]   PCR CLONING AND SEQUENCE OF 2 CDNAS ENCODING THE ALPHA-SUBUNIT AND BETA-SUBUNIT OF RABBIT CASEIN KINASE-II [J].
GUPTA, SK ;
SINGH, JP .
GENE, 1993, 124 (02) :287-290
[10]  
HATAWAY GM, 1981, J BIOL CHEM, V256, P11442