title: Osmolyte-induced folding enhances tryptic enzyme activity.
abstract: Osmolytes form a class of naturally occurring small compounds known to protect proteins in their native folded and functional states. Among the osmolytes, trimethylamine-N-oxide (TMAO) has received special interest lately because it has shown an extraordinary capability to support folding of denatured to native-like species, which show significant functional activity. Most enzymes and/or proteins are commonly stored in glycerol to maintain their activity/function. In the present study, we tested whether TMAO can be a better solute than glycerol for two commonly used proteases, trypsin and chymotrypsin. Our enzyme kinetic data suggest that the enzyme activity of trypsin is significantly enhanced in TMAO compared to glycerol, whereas chymotrypsin activity is not significantly changed in either case. These results are in accordance with the osmolyte effects on the folding of these enzymes, as judged by data from fluorescence emission spectroscopy. These results suggest that TMAO may be a better solute than glycerol to maintain optimal tryptic enzyme activity.
authors.affiliation: Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, TX 77555-1068, USA.
authors.forename: Raj
authors.initials: R
authors.lastname: Kumar
authors.forename: Justin M
authors.initials: JM
authors.lastname: Serrette
authors.forename: E Brad
authors.initials: EB
authors.lastname: Thompson
mesh_terms: D002384:Catalysis; D002918:Chymotrypsin / Q000187:drug effects / Q000378:metabolism; D004789:Enzyme Activation; D005990:Glycerol / Q000737:chemistry / Q000494:pharmacology; D007700:Kinetics; D008744:Methylamines / Q000737:chemistry / Q000494:pharmacology*; D009997:Osmotic Pressure; D011489:Protein Denaturation; D017510:Protein Folding; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D014357:Trypsin / Q000187:drug effects / Q000378:metabolism*
chemical_list: D008744:Methylamines; D002918:Chymotrypsin; D014357:Trypsin; C005855:trimethyloxamine; D005990:Glycerol