Archaeal elongation factor 1alpha from Sulfolobus solfataricus interacts with the eubacterial antibiotic GE2270A.
The thiazolyl-peptide antibiotic GE2270A, an inhibitor of the elongation factor Tu from Escherichia coli (EcEF-Tu), was used to study the effects produced in the biochemical properties of the archaeal functional analogue elongation factor 1alpha from Sulfolobus solfataricus (SsEF-1alpha). GE2270A did not substantially affect the poly(U)-directed-polyPhe incorporation catalyzed by SsEF-1alpha and the formation of the ternary complex SsEF-1alpha.GTP.Phe-tRNAPhe. On the other hand, the antibiotic was able to increase the GDP/GTP exchange rate of SsEF-1alpha; nevertheless, this improvement was not associated with an increase in the catalytic activity of the enzyme. In fact, GE2270A inhibited both the intrinsic GTPase of SsEF-1alpha (GTPaseNa) and that stimulated by ribosomes. Interestingly, GTPaseNa of both intact and C-terminal-deleted SsEF-1alpha resulted in a greater sensitivity to the antibiotic with respect to SsEF-1alpha lacking both the M- and C-terminal domains. This result suggested that, similar to what is found for EcEF-Tu, the M domain of SsEF-1alpha is the region of the enzyme most responsible for the interaction with GE2270A. The different behavior observed in the inhibition of protein synthesis with respect to EcEF-Tu can be ascribed to the different adaptive structural changes that have occurred in SsEF-1alpha during evolution.
Masullo
Mariorosario
M
Dipartimento di Scienze Farmacobiologiche, Università degli Studi di Catanzaro Magna Graecia, Roccelletta di Borgia, 88021 Catanzaro, Italy.
Cantiello
Piergiuseppe
P
Arcari
Paolo
P
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism; D006150:Guanine Nucleotides / Q000378:metabolism; D020648:Peptide Elongation Factor 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010444:Peptide Elongation Factor Tu / Q000037:antagonists & inhibitors; D010456:Peptides, Cyclic / Q000494:pharmacology*; D015202:Protein Engineering; D012329:RNA, Bacterial / Q000378:metabolism; D012360:RNA, Transfer, Phe / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D048229:Sulfolobus solfataricus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D013844:Thiazoles / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D019843:Archaeal Proteins; D006150:Guanine Nucleotides; D020648:Peptide Elongation Factor 1; D010456:Peptides, Cyclic; D012329:RNA, Bacterial; D012360:RNA, Transfer, Phe; D011994:Recombinant Proteins; D013844:Thiazoles; D010444:Peptide Elongation Factor Tu; C068122:GE 2270 A