AI Summary & Key Takeaways
AI InsightThe overall purification was approximately 105.74-fold for hCA-I and 756.64- fold for hCA-II.
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Abstract
In vitro effects of some purine analogues on human carbonic anhydrase (CA, EC 4.2.1.1) were investigated in this study. Human erythrocyte CA-I and CA-II isozymes were purified using Sepharose-4B-aniline-sulfanilamide affinity gel chromatography. Specific activity and yields were determined as 904.08 EU $mg^ {-1}$, 59.86%; 6469.23 EU $mg^ {-1}$, 56.82%, respectively. The overall purification was approximately 105.74-fold for hCA-I and 756.64- fold for hCA-II. The inhibitory effects of different nucleoside analogues on CA activity were determined at low concentrations using the esterase method under in vitro conditions. I50 values for diprophylline, acyclovir, proxyphylline, aminophylline and caffeine were calculated from activity % -[I] as 4.12 μM, 5.58 μM, 9.96 μM, 19.17 mM, and 23.21 mM for hCA-I and 1.19 μM, 2.57 μM, 4.29 μM, 4.71 mM and 4.94 mM for hCA-II respectively. Additonally, the Lineweaver-Burk curves obtained were used for the determination of $K _i$ and the inhibitor type for diprophylline, acyclovir and poxyphylline. Abbreviations: CA, carbonic anhydrase; hCA-I, human carbonic anhydrase I; hCA-II, human carbonic anhydrase II.