Location of cysteine and cystine residues in S-ribonucleases associated with gametophytic self-incompatibility

被引:31
作者
Ishimizu, T
Norioka, S
Kanai, M
Clarke, AE
Sakiyama, F
机构
[1] OSAKA UNIV,INST PROT RES,DIV PROT CHEM,SUITA,OSAKA 565,JAPAN
[2] THERMOQUEST CO LTD,TOKYO,JAPAN
[3] UNIV MELBOURNE,SCH BOT,PLANT CELL BIOL RES CTR,PARKVILLE,VIC 3052,AUSTRALIA
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1996年 / 242卷 / 03期
关键词
cysteine; disulfide bond; pyridylethylation; self-incompatibility; ribonuclease;
D O I
10.1111/j.1432-1033.1996.0627r.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
S-Ribonucleases (S-RNases) that cosegregate with S-alleles in the styles of solanaceous and rosaceous plants are associated with gametophytic self-incompatibility (GSI). The amino acid sequences of many S-RNases have been derived from cDNA sequences, but the state of half-cystines has not been clarified. We report the locations of the two free cysteine residues and four disulfide bridges of tobacco S-6-RNase and of the four disulfide bridges of Japanese pear S-4-RNase. The protein was first S-pyridylethylated at a low pH to selectively modify the free cysteines without thiol-disulfide exchange. The S-pyridylethylated protein (PE-protein) was digested with Achromobacter protease I (API) at pH 6.5 then analyzed by liquid chromatography/electrospray-ionization mass spectrometry (LC/ESI-MS). This analysis showed that tobacco S-6-RNase has two free cysteine residues, Cys77 and Cys95, and four disulfide bonds at Cys16-Cys21, Cys45-Cys94, Cys153-Cys182 and Cys165-Cys176. Similarly, four disulfide bonds were identified for pear S-4-RNase, which bears no free cysteine, at Cys15-Cys22, Cys48-Cys92, Cys156-Cys195 and Cys172-Cys183. The eight cysteines forming these four disulfide bonds are conserved in all the known S-RNases, indicative that these cross-links are important in stabilizing the tertiary structures of the self-incompatibility-associated glycoproteins in the solanaceous and rosaceous families. The LC/ESI-MS analysis also provided some structural informations regarding sugar chains attached to the S-RNases.
引用
收藏
页码:627 / 635
页数:9
相关论文
共 27 条
[1]   SEQUENCE VARIABILITY OF 3 ALLELES OF THE SELF-INCOMPATIBILITY GENE OF NICOTIANA-ALATA [J].
ANDERSON, MA ;
MCFADDEN, GI ;
BERNATZKY, R ;
ATKINSON, A ;
ORPIN, T ;
DEDMAN, H ;
TREGEAR, G ;
FERNLEY, R ;
CLARKE, AE .
PLANT CELL, 1989, 1 (05) :483-491
[2]   THE ARABIDOPSIS RIBONUCLEASE GENE RNS1 IS TIGHTLY CONTROLLED IN RESPONSE TO PHOSPHATE LIMITATION [J].
BARIOLA, PA ;
HOWARD, CJ ;
TAYLOR, CB ;
VERBURG, MT ;
JAGLAN, VD ;
GREEN, PJ .
PLANT JOURNAL, 1994, 6 (05) :673-685
[3]  
CRESTFIELD AM, 1963, J BIOL CHEM, V238, P622
[4]  
de Nettancourt D., 1977, INCOMPATIBILITY ANGI
[5]   TISSUE SULFHYDRYL GROUPS [J].
ELLMAN, GL .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1959, 82 (01) :70-77
[6]  
FRIEDMAN M, 1970, J BIOL CHEM, V245, P3868
[7]   THE RIBONUCLEASES OF HIGHER-PLANTS [J].
GREEN, PJ .
ANNUAL REVIEW OF PLANT PHYSIOLOGY AND PLANT MOLECULAR BIOLOGY, 1994, 45 :421-445
[8]   PRIMARY STRUCTURE OF A BASE NON-SPECIFIC RIBONUCLEASE FROM RHIZOPUS-NIVEUS [J].
HORIUCHI, H ;
YANAI, K ;
TAKAGI, M ;
YANO, K ;
WAKABAYASHI, E ;
SANDA, A ;
MINE, S ;
OHGI, K ;
IRIE, M .
JOURNAL OF BIOCHEMISTRY, 1988, 103 (03) :408-418
[9]  
IOERGER TR, 1991, SEX PLANT REPROD, V4, P81
[10]  
Ishimizu T, 1996, J BIOCHEM-TOKYO, V120, P326, DOI 10.1093/oxfordjournals.jbchem.a021417