The chemical modification of alpha-chymotrypsin with both hydrophobic and hydrophilic compounds stabilizes the enzyme against denaturation in water-organic media.
We considered alpha-chymotrypsin (CT) in homogeneous water-organic media as a model system to examine the influence of enzyme chemical modification with hydrophilic and hydrophobic substances on its stability, activity and structure. Both types of modifying agents may lead to considerable stabilization of the enzyme in water-ethanol and water-DMF mixtures: (i) the range of organic cosolvent concentration at which enzyme activity (Vm) is at least 100% of its initial value is broadened and (ii) the range of organic cosolvent concentration at which the residual enzyme activity is observed is increased. We found that for both types of modification the stabilization effect can be correlated with the changes in protein surface hydrophobicity/hydrophilicity brought about by the modification. Circular dichroism studies indicated that the effects of these two types of modification on CT structure and its behavior in water-ethanol mixtures are different. Differential scanning calorimetry studies revealed that after modification two or three fractions or domains, differing in their stability, can be resolved. The least stable fractions (or domains) have properties similar to native CT.
Vinogradov
A A
AA
A.N. Nesmeyanov Institute of Organoelement Compounds, 28 Vavilov str., 119991 Moscow, Russia. alvin@ineos.ac.ru
Kudryashova
E V
EV
Grinberg
V Y
VY
Grinberg
N V
NV
Burova
T V
TV
Levashov
A V
AV
D000107:Acetylation; D000478:Alkylation; D000595:Amino Acid Sequence; D000812:Anhydrides / Q000737:chemistry; D002152:Calorimetry, Differential Scanning; D002384:Catalysis; D002918:Chymotrypsin / Q000737:chemistry* / Q000378:metabolism*; D002942:Circular Dichroism; D004795:Enzyme Stability; D000431:Ethanol / Q000737:chemistry / Q000494:pharmacology; D057927:Hydrophobic and Hydrophilic Interactions; D009930:Organic Chemicals / Q000737:chemistry / Q000494:pharmacology; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D012997:Solvents / Q000737:chemistry; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D013696:Temperature; D013816:Thermodynamics; D014301:Trinitrobenzenes / Q000378:metabolism; D014867:Water / Q000737:chemistry
D000812:Anhydrides; D009930:Organic Chemicals; D012997:Solvents; D014301:Trinitrobenzenes; D014867:Water; D000431:Ethanol; D002918:Chymotrypsin; C117608:alpha-chymotrypsin