A new manner of reporting pressure results after glaucoma surgery.

PURPOSE: To evaluate to what extent contemporary glaucoma abstracts offer complete information and to suggest a new manner of pressure results reporting. MATERIALS, METHODS, AND RESULTS: Most of the 36 relevant surgical glaucoma abstracts found in one issue of International Glaucoma Review contain insufficient data-supported statements. Such abstracts cannot offer a clear picture of the study essence if economic, linguistic, or political barriers prevent access to the full text. In order to enrich abstract content and to avoid typographic space waste, a formula is suggested to provide, in one single line of symbols and figures, all the necessary data for statistical interpretation at two evolution moments: the first significative control (6 months) and the final one. CONCLUSION: The current manner of results reporting in surgical glaucoma abstracts is subject to too little standardization, allowing insufficiently data-supported statements. Abstracts, especially those printed in small-circulation language journals, should be conceived and standardized in such a manner that any abstract review reader is capable of grasping the essence of the study at first glance. The suggested manner of reporting results would bring satisfaction to all areas of the process. Publishers would save typographic space, readers would find all the necessary data for statistical analysis and comparison with other studies, and authors would be convinced that the essence of their work would penetrate in spite of any economic, linguistic, or political barriers.

A central interdomain protein joint in elongation factor G regulates antibiotic sensitivity, GTP hydrolysis, and ribosome translocation.

The antibiotic fusidic acid potently inhibits bacterial translation (and cellular growth) by lodging between domains I and III of elongation factor G (EF-G) and preventing release of EF-G from the ribosome. We examined the functions of key amino acid residues near the active site of EF-G that interact with fusidic acid and regulate hydrolysis of GTP. Alanine mutants of these residues spontaneously hydrolyzed GTP in solution, bypassing the normal activating role of the ribosome. A conserved phenylalanine in the switch II element of EF-G was important for suppressing GTP hydrolysis in solution and critical for catalyzing translocation of the ribosome along mRNA. These experimental results reveal the multipurpose roles of an interdomain joint in the heart of an essential translation factor that can both promote and inhibit bacterial translation.

Intramolecular movements in EF-G, trapped at different stages in its GTP hydrolytic cycle, probed by FRET.

Elongation factor G (EF-G) is one of several GTP hydrolytic proteins (GTPases) that cycles repeatedly on and off the ribosome during protein synthesis in bacterial cells. In the functional cycle of EF-G, hydrolysis of guanosine 5'-triphosphate (GTP) is coupled to tRNA-mRNA translocation in ribosomes. GTP hydrolysis induces conformational rearrangements in two switch elements in the G domain of EF-G and other GTPases. These switch elements are thought to initiate the cascade of events that lead to translocation and EF-G cycling between ribosomes. To further define the coupling mechanism, we developed a new fluorescent approach that can detect intramolecular movements in EF-G. We attached a fluorescent probe to the switch I element (sw1) of Escherichia coli EF-G. We monitored the position of the sw1 probe, relative to another fluorescent probe anchored to the GTP substrate or product, by measuring the distance-dependent, Förster resonance energy transfer between the two probes. By analyzing EF-G trapped at five different functional states in its cycle, we could infer the cyclical movements of sw1 within EF-G. Our results provide evidence for conformational changes in sw1, which help to drive the unidirectional EF-G cycle during protein synthesis. More generally, our approach might also serve to define the conformational dynamics of other GTPases with their cellular receptors.

Conformational changes in switch I of EF-G drive its directional cycling on and off the ribosome.

We have trapped elongation factor G (EF-G) from Escherichia coli in six, functionally defined states, representing intermediates in its unidirectional catalytic cycle, which couples GTP hydrolysis to tRNA-mRNA translocation in the ribosome. By probing EF-G with trypsin in each state, we identified a substantial conformational change involving its conserved switch I (sw1) element, which contacts the GTP substrate. By attaching FeBABE (a hydroxyl radical generating probe) to sw1, we could monitor sw1 movement (by approximately 20 A), relative to the 70S ribosome, during the EF-G cycle. In free EF-G, sw1 is disordered, particularly in GDP-bound and nucleotide-free states. On EF-G*GTP binding to the ribosome, sw1 becomes structured and tucked inside the ribosome, thereby locking GTP onto EF-G. After hydrolysis and translocation, sw1 flips out from the ribosome, greatly accelerating release of GDP and EF-G from the ribosome. Collectively, our results support a central role of sw1 in driving the EF-G cycle during protein synthesis.