Comparing the human and schistosomal hypoxanthine-guanine phosphoribosyltransferases by circular dichroism.
The hypoxanthine-guanine phosphoribosyltransferases (HGPRTases) of human and the parasitic trematode, Schistosoma mansoni, are of biomedical importance. The conformations of these two enzymes were studied by circular dichroism (CD). The schistosomal HGPRTase is estimated to contain 27% alpha-helix and 30% beta-structure. This result is consistent with what is predicted from a tertiary model (Craig, S.P., Cohen, F.E., Yuan, L., McKerrow, J.H. and Wang, C.C. (1991) in Molecular & Immunological Aspects of Parasitism (Wang, C.C., ed.), pp. 122-138, Am. Assoc. Adv. Sci., Washington DC, USA), which proposes that the enzyme is an alpha/beta barrel protein. The human enzyme is estimated to contain 21% alpha-helix and 53% beta-form. The two enzymes are different in their thermostability. The human enzyme remains active after being heated to 85 degrees C for 15 min, while the schistosomal enzyme only retains its activity at temperature below 65 degrees C. The transition temperature (T1/2) of the schistosomal HGPRTase was determined by CD measurement to be 57.5 degrees C. One of the enzyme substrates, phosphoribose pyrophosphate (PRPP), stabilizes the HGPRTases by preventing the human enzyme from unfolding at 85 degrees C and partially protecting the schistosomal enzyme from unfolding at 65 degrees C. It is suggested that the amino-acid substitutions in the human enzyme improve the spatial structure and stability of its alpha-helices, which may lead to an enhanced thermostability.
Yuan
L
L
Department of Pharmaceutical Chemistry, University of California, San Franscisco 94143.
Wu
C S
CS
Craig
S P
SP
Liu
A F
AF
Wang
C C
CC
D000595:Amino Acid Sequence; D000818:Animals; D002942:Circular Dichroism; D004795:Enzyme Stability; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000737:chemistry*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D012550:Schistosoma mansoni / Q000201:enzymology*
D011994:Recombinant Proteins; D007041:Hypoxanthine Phosphoribosyltransferase