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On the interaction of periodate oxidized GDP and its borohydride reduction product with the elongation factors Tu and G from Escherichia coli
Author(s) -
Gordon Julian,
Bodley James W.
Publication year - 1974
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(74)80026-8
Subject(s) - borohydride , periodate , elongation , escherichia coli , chemistry , philosophy , stereochemistry , materials science , biochemistry , gene , metallurgy , ultimate tensile strength , catalysis
Nothing is known about the nucleotide binding sites of polypeptide chain elongation factor EF-Tu or on the EF-G-ribosome complex (nomenclature as in ref. [ 11). Also, so far no difference has been reported between their substrate specificities (reviewed in ref. [2]). We have been investigating derivatives of GTP and GDP as an attack on both of these questions. In order to form a reactive derivative of GTP or GSP which may be useful as an active site label, we followed the strategy of Erlanger and Beiser [3]. They made an immunogenic adduct between nucleotides and serum albumin. First, they formed the dialdehyde derivative by periodated oxidation. Then they made the Schiff s base between this and the amino groups of the protein. Finally, they made a stable covalent link by borohydride reduction. With this strategy in mind, we prepared periodate oxidized GTP (GTPoX) and its borohydride reduction product (GTPoX-‘Cd). We have found that both can substitute for GDP in the binding site of the EF-G- ribosome complex, but neither can bind to EF-Tu. While we have thus far been unable to form a covalent link between GDPoX and the guanine nucleotide bind- ing site, the experiments revealed an important differ- ence between the substrate specificities of EF-Tu and EF-G. 2.