PURIFICATION AND PROPERTIES OF ZINC-METALLOPHOSPHOLIPASE-C FROM PSEUDOMONAS-FLUORESCENS

被引:11
作者
CREVEL, I [1 ]
SALLY, U [1 ]
CARNE, A [1 ]
KATAN, M [1 ]
机构
[1] INST CANC RES, CHESTER BEATTY LABS, LONDON SW3 6JB, ENGLAND
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1994年 / 224卷 / 03期
关键词
D O I
10.1111/j.1432-1033.1994.00845.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
Phospholipase C produced by Pseudomonas fluorescens, isolated as a laboratory contaminant, has been purified to apparent homogeneity by ammonium sulphate fractionation, anion-exchange and size-exclusion chromatographies. The apparent molecular mass of the purified polypeptide was 39.5 kDa. Purified preparations of phospholipase C were used to characterize its enzymic properties and to obtain amino acid sequence of the N-terminus of the molecule. The P. fluorescens phospholipase C hydrolysed PtdEtn, PtdCho and PtdSer (PtdEtn > PtdCho much greater than PtdSer) and was relatively thermostable. The enzyme was inactivated in the presence of chelating agent o-phenanthroline and the activity restored after addition of zinc. Properties of this enzyme and in particular the requirement for zinc ions for the activity, revealed similarity with the well characterised Bacillus cereus phospholipase C. Similarities with other bacterial and mammalian enzymes reported to be related to the B. cereus type are discussed.
引用
收藏
页码:845 / 852
页数:8
相关论文
共 30 条
[1]   INOSITOL TRISPHOSPHATE AND CALCIUM SIGNALING [J].
BERRIDGE, MJ .
NATURE, 1993, 361 (6410) :315-325
[2]  
BHAMIDIPATI SP, 1993, J BIOL CHEM, V268, P2431
[3]   THE REGULATION AND CELLULAR FUNCTIONS OF PHOSPHATIDYLCHOLINE HYDROLYSIS [J].
BILLAH, MM ;
ANTHES, JC .
BIOCHEMICAL JOURNAL, 1990, 269 (02) :281-291
[4]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[5]   HYDROLYSIS OF PHOSPHATIDYLCHOLINE COUPLES RAS TO ACTIVATION OF RAF PROTEIN-KINASE DURING MITOGENIC SIGNAL-TRANSDUCTION [J].
CAI, H ;
ERHARDT, P ;
TROPPMAIR, J ;
DIAZMECO, MT ;
SITHANANDAM, G ;
RAPP, UR ;
MOSCAT, J ;
COOPER, GM .
MOLECULAR AND CELLULAR BIOLOGY, 1993, 13 (12) :7645-7651
[6]   ANTIBODIES PREPARED TO BACILLUS-CEREUS PHOSPHOLIPASE-C CROSS-REACT WITH A PHOSPHATIDYLCHOLINE PREFERRING PHOSPHOLIPASE-C IN MAMMALIAN-CELLS [J].
CLARK, MA ;
SHORR, RGL ;
BOMALSKI, JS .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1986, 140 (01) :114-119
[7]  
DEHERREROS AG, 1991, J BIOL CHEM, V266, P6825
[8]  
DOI O, 1971, BIOCHIM BIOPHYS ACTA, V248, P234, DOI 10.1016/0005-2760(71)90011-7
[9]  
EXTON JH, 1990, J BIOL CHEM, V265, P1
[10]   A STUDY OF THE INCIDENCE OF DIFFERENT FLUORESCENT PSEUDOMONAS SPECIES AND BIOVARS IN THE MICROFLORA OF FRESH AND SPOILED MEAT AND FISH, RAW-MILK, CHEESE, SOIL AND WATER [J].
GENNARI, M ;
DRAGOTTO, F .
JOURNAL OF APPLIED BACTERIOLOGY, 1992, 72 (04) :281-288