OVERPRODUCTION OF THE THERMUS-THERMOPHILUS ELONGATION FACTOR-TU IN ESCHERICHIA-COLI

被引:21
作者
AHMADIAN, MR
KREUTZER, R
SPRINZL, M
机构
[1] UNIV BAYREUTH, BIOCHEM LAB, POSTFACH 101251, W-8580 BAYREUTH, GERMANY
[2] UNIV BAYREUTH, BAYREUTHER INST MAKROMOLEK FORSCH, W-8580 BAYREUTH, GERMANY
关键词
THERMUS-THERMOPHILUS; SITE-DIRECTED MUTAGENESIS; ELONGATION FACTOR-TU; ESCHERICHIA-COLI; GENE EXPRESSION; THERMOPHILIC PROTEINS;
D O I
10.1016/0300-9084(91)90145-Q
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 elongation factor Tu (EF-Tu) encoded by the tuf1 gene of the extreme thermophilic bacterium Thermus thermophilus HB8 was expressed under control of the tac promoter from the recombinant plasmid pEFTu-10 in Escherichia coli. Thermophilic EF-Tu.GDP, which amounts to as much as 35% of the cellular protein content, was separated from the E coli EF-Tu.GDP by thermal denaturation at 60-degrees-C. The overproduced E coli-born T thermophilus EF-Tu was characterized by: i) recognition through T thermophilus anti-EF-Tu antibodies; ii) analysis of the peptides obtained by cyanogen bromide cleavage; iii) thermostability; iv) guanine nucleotide binding activity in the absence and the presence of elongation factor Ts; and v) ternary complex formation with phenylalanyl-tRNA(Phe) and GTP.
引用
收藏
页码:1037 / 1043
页数:7
相关论文
共 33 条
  • [1] STUDIES ON POLYPEPTIDE-CHAIN-ELONGATION FACTORS FROM AN EXTREME THERMOPHILE, THERMUS-THERMOPHILUS-HB8 .2. CATALYTIC PROPERTIES
    ARAI, KI
    ARAI, N
    NAKAMURA, S
    OSHIMA, T
    KAZIRO, Y
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1978, 92 (02): : 521 - 531
  • [2] ARAI KI, 1972, J BIOL CHEM, V247, P7029
  • [3] STUDIES ON POLYPEPTIDE-CHAIN-ELONGATION FACTORS FROM AN EXTREME THERMOPHILE, THERMUS-THERMOPHILUS-HB8 .1. PURIFICATION AND SOME PROPERTIES OF THE PURIFIED FACTORS
    ARAI, KI
    OTA, Y
    ARAI, N
    NAKAMURA, S
    HENNEKE, C
    OSHIMA, T
    KAZIRO, Y
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1978, 92 (02): : 509 - 519
  • [4] RNA REPLICATION - FUNCTION AND STRUCTURE OF QBETA-REPLICASE
    BLUMENTHAL, T
    CARMICHAEL, GG
    [J]. ANNUAL REVIEW OF BIOCHEMISTRY, 1979, 48 : 525 - 548
  • [5] SPECIFIC ALTERATIONS OF THE EF-TU POLYPEPTIDE-CHAIN CONSIDERED IN THE LIGHT OF ITS 3-DIMENSIONAL STRUCTURE
    DUISTERWINKEL, FJ
    KRAAL, B
    DEGRAAF, JM
    TALENS, A
    BOSCH, L
    SWART, GWM
    PARMEGGIANI, A
    LACOUR, TFM
    NYBORG, J
    CLARK, BFC
    [J]. EMBO JOURNAL, 1984, 3 (01) : 113 - 120
  • [6] SPECTROPHOTOMETRIC DETERMINATION OF PROTEIN CONCENTRATION IN CELL EXTRACTS CONTAINING TRANSFER-RNA AND RIBOSOMAL-RNAS
    EHRESMAN.B
    IMBAULT, P
    WEIL, JH
    [J]. ANALYTICAL BIOCHEMISTRY, 1973, 54 (02) : 454 - 463
  • [7] G-PROTEINS - TRANSDUCERS OF RECEPTOR-GENERATED SIGNALS
    GILMAN, AG
    [J]. ANNUAL REVIEW OF BIOCHEMISTRY, 1987, 56 : 615 - 649
  • [8] PROPERTIES OF NATIVE AND NICKED ELONGATION FACTOR-TU FROM THERMUS-THERMOPHILUS-HB 8
    GULEWICZ, K
    FAULHAMMER, HG
    SPRINZL, M
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1981, 121 (01): : 155 - 162
  • [9] ROLE OF GUANOSINE 5'-TRIPHOSPHATE IN POLYPEPTIDE-CHAIN ELONGATION
    KAZIRO, Y
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA, 1978, 505 (01) : 95 - 127
  • [10] MOLECULAR-CLONING AND SEQUENCE DETERMINATION OF THE TUF GENE CODING FOR THE ELONGATION-FACTOR TU OF THERMUS-THERMOPHILUS HB8
    KUSHIRO, A
    SHIMIZU, M
    TOMITA, K
    [J]. EUROPEAN JOURNAL OF BIOCHEMISTRY, 1987, 170 (1-2): : 93 - 98