Amino acid sequence of elongation factor Tu. Characterization and alignment of the cyanogen bromide fragments and location of the cysteine residues

Laursen, Richard A.; Nagarkatti, S; Miller, David L.

doi:10.1016/0014-5793(77)80416-x

Cited by 25 publications

(11 citation statements)

References 23 publications

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“…For example, it can give information about the role of the amino-terminal segment in the activities of the factor related to protein synthesis as well as for alternative functions. In particular, it has recently been suggested [29] that preliminary sequence data are consistent with the observation of Arai eta/. [7] that the first intermediate of EF-Tu was partially inactive, since it was found that the cysteine responsible for the binding of aminoacyl tRNA [30] is located near the amino terminus of the protein.…”

Section: Discussionsupporting

confidence: 77%

Limited Proteolysis of Elongation Factor Tu from Escherichia coli

Jacobson

Rosenbusch

1977

European Journal of Biochemistry

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Limited proteolysis of native elongation factor Tu (Mr 44000) by trypsin occurs in at least three distinct steps. The first intermediate arises through cleavage at a site about 65 residues from the amino‐terminal end of the protein. It is functionally active [Jacobson, G. R. & Rosenbusch, J. P. (1976) Biochemistry, 15, 5105–5110] and is partially protected from further degradation by the antibiotic kirromycin. The second step converts this intermediate to one of similar size (Mr 37000) which now is partially inactivated. It is likely to be identical with the intermediate described by Arai et al. [(1976) J. Biochem. Tokyo, 79, 69–83]. In the third step, the partially inactive intermediate is cleaved without any apparent change in the functional properties tested. The resulting two trypsin‐resistant fragments have molecular weights of 24000 and 14000, and remain associated under nondenaturing conditions. When either of these polypeptides, after isolation in 8 M urea, is allowed to renature, no significant reactivation of GDP binding is observed unless the isolated fragments are mixed before renaturation. These results show that the two fragments are structurally and functionally interdependent.

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Section: Discussionsupporting

confidence: 77%

Limited Proteolysis of Elongation Factor Tu from Escherichia coli

Jacobson

Rosenbusch

1977

European Journal of Biochemistry

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“…So far we have detected no differences in the sequences of EF-Tu isolated from these strains. Peptides were generated by cleavage with cyanogen bromide or limited trypsinolysis as described (14)(15)(16)(17); large peptides were further fragmented with trypsin, Staphylococcus aureus protease, or chymotrypsin. Amino acid sequences were determined either by the manual dansyl-Edman procedure (18)(19)(20)(21)(22)(23) or by automated solid-phase Edman degradation (24,25).…”

mentioning

confidence: 99%

Primary structure of elongation factor Tu from Escherichia coli.

Arai¹,

Clark²,

Duffy³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

182

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The amino acid sequence of elongation factor Tu (EF-Tu) from Escherichia coli has been determined. EF-Tu is a single-chain polypeptide composed of 393 amino acids (M, 43,225 for the species bearing COOH-terminal serine). The NHrterminal serine is acetylated, and lysine-56 is partially methylated. The sites of facile tryptic cleavage are at arginines 44 and 58 and at lysine-263. The cysteinyl residues associated with aminoacyl-tRNA and guanosine nucleotide binding activities are residues 81 and 137, respectively. The COOH-terminal amino acid is heterogeneous in that analyses of the COOH-terminal peptides isolated from different EF-Tu preparations gave position 393 as glycine and serine in ratios (Gly/ Ser) ranging from about 0.7 to 3.

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“…Results obtained in this laboratory (Frank and A.W., to be published) on the amino acid sequence of EF-Tu from B. stearothennophilus indicate that its 2 sulphydryl groups occupy positions in the primary structure analogous to SH 1 and SH 2 in the I?. coli factor sequence [8,9]. Thus a direct comparison of the functions of the sulphydryl groups of the two proteins is permissible.…”

Section: Discussionmentioning

confidence: 99%