HYBRID F-0 COMPLEXES OF THE ATP SYNTHASES OF SPINACH-CHLOROPLASTS AND ESCHERICHIA-COLI - IMMUNOPRECIPITATION AND MUTANT ANALYSES

被引:7
作者
BURKOVSKI, A [1 ]
DECKERSHEBESTREIT, G [1 ]
ALTENDORF, K [1 ]
机构
[1] UNIV OSNABRUCK,FACHBEREICH BIOL CHEM,MIKROBIOL ABT GRP,D-49069 OSNABRUCK,GERMANY
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1994年 / 225卷 / 03期
关键词
D O I
10.1111/j.1432-1033.1994.1221b.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
Hybrid F-0 complexes of the ATP synthases of spinach chloroplast (CF0) and Escherichia coli (EF(0)) were investigated. Immunoprecipitations with polyclonal antibodies against the different F-0 subunits clearly revealed that hybrid F-0 complexes derived from CF0 subunit III and EF(0) subunits a and b were formed in vivo. In addition, the ATPase activities of the hybrid ATP synthase, measured in everted cytoplasmic membranes of an atpE mutant strain transformed with the atpH gene coding for CF0 III, were comparable to activities obtained for the same mutant strain complemented with the atpE gene (EF(0) c). Nevertheless, CF0 III was not able to replace EF(0) c functionally, since the strain containing the hybrid ATP synthase was not able to grow on succinate. In order to investigate the reason for this lack of function, hybrid proteolipids of CF0 III and EF(0) c were constructed. Only a chimaeric protein comprising the seven N-terminal amino acid residues from CF0 III and the remaining part of EF(0) c was able to replace wild-type EF(0) c, whereas hybrid proteins with 13 and 33 N-terminal amino acids of CF0 III were not functional. The results suggested that a network of interactions between the subunits essential for proton translocation and/or coupling of the F-1 part exists, which was optimized for each species during evolution, although the overall structure of F0F1 complexes has been conserved.
引用
收藏
页码:1221 / 1228
页数:8
相关论文
共 69 条
[1]   AUTOMATED CONTINUOUS ASSAY OF MEMBRANE-BOUND AND SOLUBLE ATPASES AND RELATED ENZYMES [J].
ARNOLD, A ;
WOLF, HU ;
ACKERMANN, BP ;
BADER, H .
ANALYTICAL BIOCHEMISTRY, 1976, 71 (01) :209-213
[2]   COPPER ENZYMES IN ISOLATED CHLOROPLASTS - POLYPHENOLOXIDASE IN BETA-VULGARIS [J].
ARNON, DI .
PLANT PHYSIOLOGY, 1949, 24 (01) :1-15
[3]  
Ausubel FM., 1995, MOL REPROD DEV, V3rd edn, DOI DOI 10.1002/MRD.1080010210
[4]   LINKAGE MAP OF ESCHERICHIA-COLI K-12, EDITION-8 [J].
BACHMANN, BJ .
MICROBIOLOGICAL REVIEWS, 1990, 54 (02) :130-197
[5]   RECONSTITUTION OF PHOTOPHOSPHORYLATION WITH COUPLING FACTOR-I ATPASES FROM THE THERMOPHILIC BACTERIUM PS3 AND LETTUCE CHLOROPLASTS [J].
BARZVI, D ;
YOSHIDA, M ;
SHAVIT, N .
BIOCHIMICA ET BIOPHYSICA ACTA, 1985, 806 (03) :341-347
[6]  
BRUSILOW WSA, 1989, J BIOL CHEM, V264, P1528
[7]   EXPRESSION OF SUBUNIT-III OF THE ATP SYNTHASE FROM SPINACH-CHLOROPLASTS IN ESCHERICHIA-COLI [J].
BURKOVSKI, A ;
DECKERSHEBESTREIT, G ;
ALTENDORF, K .
FEBS LETTERS, 1990, 271 (1-2) :227-230
[8]   ONE-STEP PREPARATION OF COMPETENT ESCHERICHIA-COLI - TRANSFORMATION AND STORAGE OF BACTERIAL-CELLS IN THE SAME SOLUTION [J].
CHUNG, CT ;
NIEMELA, SL ;
MILLER, RH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (07) :2172-2175
[10]   THE ORGANIZATION AND SEQUENCE OF THE GENES FOR ATP SYNTHASE SUBUNITS IN THE CYANOBACTERIUM SYNECHOCOCCUS-6301 - SUPPORT FOR AN ENDOSYMBIOTIC ORIGIN OF CHLOROPLASTS [J].
COZENS, AL ;
WALKER, JE .
JOURNAL OF MOLECULAR BIOLOGY, 1987, 194 (03) :359-383