Identification of highly methylated arginine residues in an endogenous 20-kDa polypeptide in cancer cells

被引:15
作者
Gu, HM
Park, SH
Park, GH
Lim, IK
Lee, HW
Paik, WK
Kim, S [1 ]
机构
[1] Korea Univ, Sch Med, Dept Biochem, Seoul 136701, South Korea
[2] Korea Univ, Grad Sch Biotechnol, Seoul 136701, South Korea
[3] Ajou Univ, Sch Med, Dept Biochem, Suwon 442749, South Korea
[4] Sungkyunkwan Univ, Coll Pharm, Suwon 440746, South Korea
关键词
protein-arginine methylation; 20-kDa polypeptide; cell proliferation;
D O I
10.1016/S0024-3205(99)00300-8
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
Enzymatic methylation of endogenous proteins in several cancer cell lines was investigated to understand a possible relationship between protein-arginine methylation and cellular proliferation. Cytosolic extracts prepared from several cancer cells (HeLa, HCT-48, A549, and HepG2) and incubated with S-adenosyl-L-[methyl-H-3]methionine revealed an intensely [methyl-H-3]-labeled 20-kDa polypeptide. On the other hand, cytosolic extracts prepared from normal colon cells did not show any methylation of the 20-kDa protein under identical conditions. To identify nature of the 20-kDa polypeptide, purified histones were methylated with HCT-48 cytosolic extracts and analyzed by SDS-PAGE. However, none of the histones comigrated with the methylated 20-kDa polypeptide, indicating that it is unlikely to be any of the histone subclasses. The [methyl-H-3]group in the 20-kDa polypeptide was stable at pH 10-11 (37 degrees C for 30 min) and methylation was not stimulated by GTP gamma S (4 mM), thus the reaction is neither carboxyl methylesterification on isoaspartyl residues, nor on C-terminal farnesylated cysteine. The present study together with the previous identification of N-G-methylated arginine residues in the HCT-48 cytosol fraction suggests that this novel endogenous 20-kDa arginine-methylation is a cellular proliferation-related posttranslational modification reaction.
引用
收藏
页码:737 / 745
页数:9
相关论文
共 38 条
[1]   A protein-arginine methyltransferase binds to the intracytoplasmic domain of the IFNAR1 chain in the type I interferon receptor [J].
Abramovich, C ;
Yakobson, B ;
Chebath, J ;
Revel, M .
EMBO JOURNAL, 1997, 16 (02) :260-266
[2]   SPECIFIC ENZYMIC METHYLATION OF AN ARGININE IN EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS PROTEIN FROM HUMAN MYELIN [J].
BALDWIN, GS ;
CARNEGIE, PR .
SCIENCE, 1971, 171 (3971) :579-&
[3]   RAS GENES [J].
BARBACID, M .
ANNUAL REVIEW OF BIOCHEMISTRY, 1987, 56 :779-827
[4]   POST-SYNTHETIC MODIFICATIONS OF NUCLEAR PROTEINS HIGH MOBILITY GROUP PROTEINS ARE METHYLATED [J].
BOFFA, LC ;
STERNER, R ;
VIDALI, G ;
ALLFREY, VG .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1979, 89 (04) :1322-1327
[5]  
BORUN TW, 1972, J BIOL CHEM, V247, P4288
[6]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[7]  
BUCKLAND P, 1992, J BIOL CHEM, V267, P18432
[8]   REGULATION OF ALTERNATIVE SPLICING IN-VIVO BY OVEREXPRESSION OF ANTAGONISTIC SPLICING FACTORS [J].
CACERES, JF ;
STAMM, S ;
HELFMAN, DM ;
KRAINER, AR .
SCIENCE, 1994, 265 (5179) :1706-1709
[9]   INHIBITION OF PHOSPHOLIPID METHYLATION BY A CYTOSOLIC FACTOR [J].
CHIVA, VA ;
MATO, JM .
BIOCHEMICAL JOURNAL, 1984, 218 (02) :637-639
[10]  
DESROSIERS R, 1988, J BIOL CHEM, V263, P4686