Nucleotide and Mg2+ Induced Conformational Changes in GroEL can be Detected by Sulfhydryl Labeling

The accessibility of fluorescein-5-maleimide to sulfhydryl groups in the molecular chaperone GroEL was used to follow structural rearrangements in the protein triggered by binding Mg²⁺ and/or adenine nucleotides. Three peptides, each containing one of the cysteines of GroEL (C138, C458 and C519) were identified. GroEL labeled in 50mM TrisHCl, pH 7.8, incorporated ~0.3 labels each on C138 and C458. With 10mM MgCl₂, the labeling increased to ~0.8 labels each on C138 and C458. The increase was partially due to a conformational change which occurred upon Mg²⁺ binding as well as to an increase in ionic strength. When ADP, ATP, or AMP-PNP were added to a solution of GroEL and Mg²⁺, C138 incorporated ~0.8 labels, while C458 incorporated ~0.1 labels. These results suggest that the binding of adenine nucleotides changed the conformation of GroEL and made a previously highly exposed sulfhydryl group inaccessible. GroEL slowly dissociated into monomers when it was extensively labeled at C458. GroEL labeled with fluorescein-5-maleimide, under any of the conditions examined, was able to bind but not release active rhodanese. The observed variations in sulfhydryl accessibility are consistent with mechanisms that suggest binding of GroES to GroEL differs from the binding of substrate protein to GroEL, and that the binding of Mg²⁺ or Mg-adenine nucleotides results in conformational changes in GroEL.     

Relative Risk of Involuntary Injuries Among Currently Employed Adults in the U.S.

It can be very informative to compare the risk of injury for different occupations, places where accidents happen, demographic variables, and other characteristics. However, most epidemiologic literature shows that multiple logistic regression or odds ratio were commonly used to compare the relationship between the variables on two or more levels. A multiple logistic regression could be used for multiple comparisons assuming that the model correctly reflects the situation at hand. But it might not be a true assumption. Assuming no model, we defined the relative risk to compare risks of injury for different occupations, and applied this method to the data collected by the National Center for Health Statistics (NCHS) National Health Interview Survey (NHIS). This application showed that the relative risk of injury varied by occupation and by place of accident, sex, race, and age.