Purification and kinetic properties of rabbit liver paraoxonase 1

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Tülin BayrakKamer Kilinç

Abstract

Paraoxonase 1 (PON1) is synthesized in the liver and secreted into the blood, where it is associated exclusively with HDL. In this study, rabbit liver PON1 enzyme was purified to homogeneity using a new purification approach, and the kinetic properties of the enzyme were investigated using phenyl acetate and homocysteine thiolactone as substrates. Rabbit liver PON1 was purified through the preparation of liver microsomal fraction, Sephacryl S300 HR gel filtration chromatography, DEAE Trisacryl M ion-exchange chromatography and hydroxyapatite chromatography steps. Using this method, rabbit liver PON1 was purified 576 times with a specific activity of 2726 U/mg protein. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis revealed the obtained enzyme as a single protein band close to 40 kDa. The Km of the this enzyme was found as 0.55+/-0.024 mM for phenyl acetate and 17.31+/-1.2 mM for homocysteine thiolactone. In this study, a new approach was used to purify PON1 enzyme from rabbit liver and for the first time in the literature, its kinetics was studied with homocysteine thiolactone as substrate.

References

Oct 22, 1991·Biochemistry·C E FurlongJ W Crabb
Mar 27, 2001·Pharmacogenetics·C E WatsonB N La Du
Jan 13, 2006·The Journal of Biological Chemistry·Olga Khersonsky, Dan S Tawfik
Sep 4, 2009·Biological Trace Element Research·Deniz Ekinci, Sükrü Beydemir

❮ Previous
Next ❯

Citations

May 31, 2012·Amino Acids·Joanna Perła-Kaján, Hieronim Jakubowski
Jul 23, 2013·Indian Journal of Endocrinology and Metabolism·Sunil K KotaKirtikumar D Modi
Nov 3, 2010·Chemical Biology & Drug Design·Deniz EkinciClaudiu T Supuran

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bioinformatics in Biomedicine

Bioinformatics in biomedicine incorporates computer science, biology, chemistry, medicine, mathematics and statistics. Discover the latest research on bioinformatics in biomedicine here.

Related Papers

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Ahmet BayrakKamer Kılınç
Clinica Chimica Acta; International Journal of Clinical Chemistry
F EndoI Matsuda
The International Journal of Biochemistry & Cell Biology
Ji Young SonIn Kook Park
© 2021 Meta ULC. All rights reserved