Purification of porcine brain protein phosphatase 2A leucine carboxyl methyltransferase and cloning of the human homologue

被引:120
作者
De Baere, I [1 ]
Derua, R [1 ]
Janssens, V [1 ]
Van Hoof, C [1 ]
Waelkens, E [1 ]
Merlevede, W [1 ]
Goris, J [1 ]
机构
[1] Katholieke Univ Leuven, Dept Geneeskunde, Afeling Biochem, B-3000 Louvain, Belgium
关键词
D O I
10.1021/bi991646a
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 carboxyl methyltransferase, which is claimed to exclusively methylate the carboxyl group of the C-terminal leucine residue of the catalytic subunit of protein phosphatase 2A (Leu(309)), was purified from porcine brain. On the basis of tryptic peptides, the cDNA encoding the human homologue was cloned. The cDNA of this gene encodes for a protein of 334 amino acids with a calculated M-r of 38 305 and a predicted pi of 5.72. Database screening reveals the presence of this protein in diverse phyla. Sequence analysis shows that the novel methyltransferase is distinct from other known protein methyltransferases, sharing only sequence motifs supposedly involved in the binding of adenosylmethionine. The recombinant protein expressed in bacteria is soluble and the biophysical, catalytic, and immunological properties are indistinguishable from the native enzyme, The methylation of PP2A by the recombinant protein is restricted to Leu(309) of PP2A(C). No direct effects on phosphatase activity changes were observed upon methylation of the dimeric or trimeric forms of PP2A.
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页码:16539 / 16547
页数:9
相关论文
共 44 条
[1]   Protein methylation: a signal event in post-translational modification [J].
Aleta, JM ;
Cimato, TR ;
Ettinger, MJ .
TRENDS IN BIOCHEMICAL SCIENCES, 1998, 23 (03) :89-91
[2]   Gapped BLAST and PSI-BLAST: a new generation of protein database search programs [J].
Altschul, SF ;
Madden, TL ;
Schaffer, AA ;
Zhang, JH ;
Zhang, Z ;
Miller, W ;
Lipman, DJ .
NUCLEIC ACIDS RESEARCH, 1997, 25 (17) :3389-3402
[3]  
[Anonymous], 1988, Antibodies: A Laboratory Manual
[4]   PURIFICATION AND CHARACTERIZATION OF 2 DISTINCT ISOZYMES OF PROTEIN CARBOXYMETHYLASE FROM BOVINE BRAIN [J].
ASWAD, DW ;
DEIGHT, EA .
JOURNAL OF NEUROCHEMISTRY, 1983, 40 (06) :1718-1726
[5]   Methylated C-terminal leucine residue of PP2A catalytic subunit is important for binding of regulatory Bα subunit [J].
Bryant, JC ;
Westphal, RS ;
Wadzinski, BE .
BIOCHEMICAL JOURNAL, 1999, 339 :241-246
[6]  
CAYLA X, 1994, J BIOL CHEM, V269, P15668
[7]  
CHEN J, 1994, J BIOL CHEM, V269, P7957
[8]   REGULATION OF PROTEIN SERINE-THREONINE PHOSPHATASE TYPE-2A BY TYROSINE PHOSPHORYLATION [J].
CHEN, J ;
MARTIN, BL ;
BRAUTIGAN, DL .
SCIENCE, 1992, 257 (5074) :1261-1264
[9]  
CLARKE S, 1985, ANNU REV BIOCHEM, V54, P479, DOI 10.1146/annurev.biochem.54.1.479
[10]   MULTIPLE SEQUENCE ALIGNMENT WITH HIERARCHICAL-CLUSTERING [J].
CORPET, F .
NUCLEIC ACIDS RESEARCH, 1988, 16 (22) :10881-10890