ARGININE DECARBOXYLASE OF OATS IS CLIPPED FROM A PRECURSOR INTO 2-POLYPEPTIDES FOUND IN THE SOLUBLE ENZYME

被引:29
作者
MALMBERG, RL
SMITH, KE
BELL, E
CELLINO, ML
机构
[1] Botany Department, University of Georgia, Athens
[2] Department of Biochemistry, Texas A and M University, College Station
关键词
D O I
10.1104/pp.100.1.146
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
We have examined soluble oat (Avena sativa) arginine decarboxylase by probing its structure with polyclonal antibodies that separately recognize amino-terminal and carboxyl-terminal antigens and with a monoclonal antibody that immunoprecipitates enzyme activity. These experiments indicated that oat arginine decarboxylase is clipped from a 66,000-D precursor polypeptide into 42,000- and 24,000-D produce polypeptides. Both of these are found in the enzyme and may be held together by disulfide bonds. A full-length precursor protein could not be detected in plants but could be produced by expression of the cDNA in Escherichia coli. Analysis of the expression of the cDNA in E. coli, with antibodies and using pulse labeling with [S-35]methionine, indicated that the bulk of the expressed protein was the full-length 66,000-D form. Small amounts of 42,000- and 24,000-D polypeptides could also be detected. A reconstruction experiment, adding a radioactively labeled full-length protein isolated from E. coli to powdered oat leaves, supported the idea that the protein extraction method used for western blots was not likely to result in artifactual proteolytic degradation.
引用
收藏
页码:146 / 152
页数:7
相关论文
共 20 条
[1]   ANALYSIS OF A CDNA-ENCODING ARGININE DECARBOXYLASE FROM OAT REVEALS SIMILARITY TO THE ESCHERICHIA-COLI ARGININE DECARBOXYLASE AND EVIDENCE OF PROTEIN PROCESSING [J].
BELL, E ;
MALMBERG, RL .
MOLECULAR & GENERAL GENETICS, 1990, 224 (03) :431-436
[2]   CATALYTIC IRREVERSIBLE INHIBITION OF BACTERIAL AND PLANT ARGININE DECARBOXYLASE ACTIVITIES BY NOVEL SUBSTRATE AND PRODUCT ANALOGS [J].
BITONTI, AJ ;
CASARA, PJ ;
MCCANN, PP ;
BEY, P .
BIOCHEMICAL JOURNAL, 1987, 242 (01) :69-74
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   BIOSYNTHETIC ARGININE DECARBOXYLASE IN ESCHERICHIA-COLI IS SYNTHESIZED AS A PRECURSOR AND LOCATED IN THE CELL-ENVELOPE [J].
BUCH, JK ;
BOYLE, SM .
JOURNAL OF BACTERIOLOGY, 1985, 163 (02) :522-527
[5]   MONOCLONAL-ANTIBODIES WITH DIFFERING AFFINITIES TO THE RED-ABSORBING AND FAR-RED-ABSORBING FORMS OF PHYTOCHROME [J].
CORDONNIER, MM ;
GREPPIN, H ;
PRATT, LH .
BIOCHEMISTRY, 1985, 24 (13) :3246-3253
[6]   BIOCHEMICAL DIFFERENTIATION IN THE TOBACCO FLOWER PROBED WITH MONOCLONAL-ANTIBODIES [J].
EVANS, PT ;
HOLAWAY, BL ;
MALMBERG, RL .
PLANTA, 1988, 175 (02) :259-269
[7]   DO POLYAMINES HAVE ROLES IN PLANT DEVELOPMENT [J].
EVANS, PT ;
MALMBERG, RL .
ANNUAL REVIEW OF PLANT PHYSIOLOGY AND PLANT MOLECULAR BIOLOGY, 1989, 40 :235-269
[8]  
FLORES H, 1990, CURRENT TOPICS PLANT, V5
[9]  
HARLOW E, 1988, ANTIBODIES LABORATOR
[10]   UTILIZATION OF PUTRESCINE IN TOBACCO CELL-LINES RESISTANT TO INHIBITORS OF POLYAMINE SYNTHESIS [J].
HIATT, A ;
MALMBERG, RL .
PLANT PHYSIOLOGY, 1988, 86 (02) :441-446