JZUS - Journal of Zhejiang University SCIENCE

Journal of Zhejiang University SCIENCE B 2011 Vol.12 No.3 P.180-188

Kinetic analysis of γ-glutamyltransferase reaction process for measuring activity via an integration strategy at low concentrations of γ-glutamyl p-nitroaniline

Author(s): Zhi-rong Li, Yin Liu, Xiao-lan Yang, Jun Pu, Bei-zhong Liu, Yong-hua Yuan, Yan-ling Xie, Fei Liao
Affiliation(s): Key Laboratory of Medical Laboratory Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016, China, Unit for Biotransformation and Protein Biotechnology, Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University, Chongqing 400016, China
Corresponding email(s): liaofeish@yahoo.com, liaofeish@cqmu.edu.cn
Key Words: Integration strategy, Chromogenic substrate, Data processing, γ, -Glutamyltransferase, Kinetic analysis, Serum enzyme assay

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Abstract: At 0.12 mmol/L γ;-glutamyl p-nitroaniline (GGPNA), an improved integrated method was developed for kinetic analysis of γ;-Glutamyltransferase (GGT) reaction process and the integration with the classical initial rate method to measure serum GGT. For the improved integrated method, an integrated rate equation, which used the predictor variable of reaction time and considered inhibitions by both GGPNA and products, was nonlinearly fit to GGT reaction processes. For the integration strategy, classical initial rates were estimated when GGPNA consumption percentages were below 50%; otherwise, maximal reaction rates of GGT were estimated by the improved integrated method and converted into initial rates according to the differential rate equation at 0.11 mmol/L GGPNA. The integration strategy was validated using optimized GGT kinetic parameters and 10-s intervals to record reaction curves within 8.0 min. By the integration strategy, there was a linear response from 0.9 to 32.0 U/L GGT, coefficients of variation were below 3.5% for GGT from 8.0 to 32.0 U/L (n=5), and GGT activities in clinical sera responded linearly to their classical initial rates at 2.00 mmol/L GGPNA with an expected slope. Therefore, the integration strategy was successful in measuring GGT at 0.12 mmol/L GGPNA.

Reference

[1]Atkins, G.L., Nimmo, I.A., 1973. The reliability of Michaelis-Menten constants and maximum velocities estimated by using the integrated Michaelis-Menten equation. Biochem. J., 135(3):779-784.

[2]Castonguay, R., Halim, D., Morin, M., Furtos, A., Lherbet, C., Bonneil, E., Thibault, P., Keillor, J.W., 2007. Kinetic characterization and identification of the acylation and glycosylation sites of recombinant human γ-glutamyl-transpeptidase. Biochemistry, 46(43):12253-12262.

[3]Cook, N.D., Peters, T.J., 1986. The simultaneous hydrolysis of glutathione and glutamine by rat kidney γ-glutamyltransferase. Biochim. Biophys. Acta, 884(1):207-210.

[4]Cook, N.D., Upperton, K.P., Challis, B.C., Peters, T.J., 1987. The donor specificity and kinetics of the hydrolysis reaction of γ-glutamyltransferase. Biochim. Biophys. Acta, 914(3):240-245.

[5]Dvorakova, L., Krusek, J., Stastny, F., Lisy, V., 1996. Analysis of kinetic properties of γ-glutamyl transpeptidase from rat kidney. Gen. Physiol. Biophys., 15(5):403-413.

[6]Fossati, P., Melzi d′Eril, G.V., Tarenghi, G., Prencipe, L., Berti, G., 1986. A kinetic colorimetric assay of γ-glutamyl-transferase. Clin. Chem., 32(8):1581-1584.

[7]Gella, F.J., Gubern, G., Vidal, R., Canalias, F., 1997. Determination of total and pancreatic α-amylase in human serum with 2-chloro-4-nitrophenyl-α-d-maltotrioside as substrate. Clin. Chim. Acta, 259(1-2):147-160.

[8]Giral, P., Jacob, N., Dourmap, C., Hansel, B., Carrié, A., Bruckert, E., Girerd, X., Chapman, M.J., 2008. Elevated γ-glutamyltransferase activity and perturbed thiol profile are associated with features of metabolic syndrome. Arterioscler. Thromb. Vasc. Biol., 28(3):587-593.

[9]Jo, S.K., Lee, W.Y., Rhee, E.J., Won, J.C., Jung, C.H., Park, C.Y., Oh, K.W., Park, S.W., Kim, S.W., 2009. Serum γ-glutamyl transferase activity predicts future development of metabolic syndrome defined by 2 different criteria. Clin. Chim. Acta, 403(1-2):234-240.

[10]Liao, F., Liu, W.L., Zhou, Q.X., Zeng, Z.C., Zuo, Y.P., 2001. Assay of serum arylesterase activity by fitting to the reaction curve with an integrated rate equation. Clin. Chim. Acta, 314(1-2):67-76.

[11]Liao, F., Tian, K.C., Yang, X., Zhou, Q.X., Zeng, Z.C., Zuo, Y.P., 2003. Kinetic substrate quantification by nonlinear fitting reaction curve to integrated Michaelis-Menten equation. Anal. Bioanal. Chem., 375(6):756-762.

[12]Liao, F., Zhu, X.Y., Wang, Y.M., Zuo, Y.P., 2005. The comparison of the estimation of enzyme kinetic parameters by fitting reaction curve to the integrated Michaelis-Menten rate equations of different predictor variables. J. Biochem. Biophys. Methods, 62(1):13-24.

[13]Liao, F., Zhao, Y.S., Zhao, L.N., Tao, J., Zhu, X.Y., Liu, L., 2006. The evaluation of a direct kinetic method for serum uric acid assay by predicting the background absorbance of uricase reaction solution with an integrated method. J. Zhejiang Univ.-Sci. B, 7(6):497-502.

[14]Liao, F., Zhao, L.N., Zhao, Y.S., Tao, J., Zuo, Y.P., 2007. Integrated rate equation considering product inhibition and its application to kinetic assay of serum ethanol. Anal. Sci., 23(4):439-444.

[15]Liao, F., Yang, D.Y., Tang, J.Q., Yang, X.L., Liu, B.Z., Zhao, Y.S., Zhao, L.N., Liao, H., Yu, M.A., 2009. The measurement of serum cholinesterase activities by an integration strategy with expanded linear ranges and negligible substrate-activation. Clin. Biochem., 42(9):926-928.

[16]Lim, J.S., Lee, D.H., Park, J.Y., Jin, S.H., Jacobs, D.R., 2007. A strong interaction between serum γ-glutamyltransferase and obesity on the risk of prevalent type 2 diabetes: results from the third national health and nutrition examination survey. Clin. Chem., 53(6):1092-1098.

[17]Liu, B.Z., Zhao, Y.S., Zhao, L.N., Xie, Y.L., Zhu, S., Li, Z.R., Liu, Y., Lu, W., Yang, X.L., Xie, G.M., et al., 2009. An integration strategy to estimate the initial rates of enzyme reactions with much expanded linear ranges using uricases as models. Anal. Chim. Acta, 631(1):22-28.

[18]London, J.W., Shaw, L.M., Fetterolf, D., Garfinkel, D., 1976. Determination of the mechanism and kinetic constants for hog kidney γ-glutamyltransferase. Biochem. J., 157(3):609-617.

[19]Newman, P.F.J., Atkins, G.L., Nimmo, I.A., 1974. The effects of systematic error on the accuracy of Michaelis constant and maximum velocities estimated by using the integrated Michaelis-Menten equation. Biochem. J., 143(3):779-781.

[20]Orsi, B.A., Tipton, K.F., 1979. Kinetic analysis of progress curves. Methods Enzymol., 63:159-183.

[21]Ruttmann, E., Brant, L.J., Concin, H., Diem, G., Rapp, K., Ulmer, H., 2005. γ-Glutamyltransferase as a risk factor for cardiovascular disease mortality. An investigation in a cohort of 163 944 Austrian adults. Circulation, 112(14):2130-2137.

[22]Shaw, L.M., London, J.W., Fetterolf, D., Garfinkel, D., 1972. γ-Glutamyltransferase: kinetic properties and assay conditions when γ-glutamyl-4-nitroanilide and its 3-carboxy derivative are used as donor substrates. Clin. Chem., 23(1):79-85.

[23]Stein, R.L., DeCicco, C., Nelson, D., Thomas, B., 2001. Slow-binding inhibition of γ-glutamyl transpeptidase by γ-boroGlu. Biochemistry, 40(19):5804-5811.

[24]Tsao, F.H., Shanmuganayagam, D., Zachman, D.K., Khosravi, M., Folts, J.D., Meyer, K.C., 2007. A continuous fluorescence assay for the determination of calcium-dependent secretory phospholipase A2 activity in serum. Clin. Chim. Acta, 379(1-2):119-123.

[25]Turgut, O., Yilmaz, A., Yalta, K., Karadas, F., Birhan, Y.M., 2006. γ-Glutamyltransferase is a promising biomarker for cardiovascular risk. Med. Hypotheses, 67(5):1060-1064.

[26]Walsh, R., Martin, E., Darvesh, S., 2010. A method to describe enzyme-catalyzed reactions by combining steady state and time course enzyme kinetic parameters. Biochim. Biophys. Acta, 1800(1):1-5.

[27]Whitfield, J.B., 2001. γ-Glutamyl transferase. Crit. Rev. Clin. Lab. Sci., 38(4):263-355.

[28]Winn-Deen, E.S., David, H., Sigler, G., Chavez, R., 1988. Development of a direct assay for alpha-amylase. Clin. Chem., 34(10):2005-2008.

[29]Yamada, M., Fujita, T., 2007. New procedure for the measurement of pancreatic lipase activity in human serum using a thioester substrate. Clin. Chim. Acta, 383(1-2):85-90.

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