Abstract
Some molecular properties of the elongation factor Tu of protein synthesis purified in an aggregated state from gram-positiveStreptomyces aureofaciens were studied and compared with those of Tu from gram-negativeEscherichia coli. Electrofocussing under reducing conditions showed that the molecule of EF-Tu fromS. aureofaciens has an isoelectric point shifted more to the acidic side compared with EF-Tu fromE. coli. A comparison of amino acid composition revealed minor differences in the content of several amino acids in the two factors and showed that EF-Tu fromS. aureofaciens contains four half-cystines per molecule. Under denaturing conditions only two mercapto groups reacted with 5,5’-dithiobis(2-nitrobenzoic acid). Limited tryptic digestion of aggregated EF-Tu fromS. aureofaciens yields six fragments: the four main fragments are of a similar size as those of theE. coli factor. All fragments detected after trypsin digestion ofS. aureofaciens EF-Tu were immunologically cross-reactive with antibodies againstE. coli EF-Tu. However, even after 2 h of the reaction there still remains a small part of streptomycete factor uncleaved, which documents high resistance of aggregated EF-Tu towards trypsin.
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We are indebted to Drs. J. Jonák, J. Pohl and V. Hořejší for helpful advices and discussions and to Dr. K. Mikulík for providing us with TPCK-treated trypsin.
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Weiser, J., Šebo, P. Molecular properties of elongation factor Tu fromStreptomyces aureofaciens andEscherichia coli . Folia Microbiol 33, 81–87 (1988). https://doi.org/10.1007/BF02928072
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DOI: https://doi.org/10.1007/BF02928072