Single chain antibody displays glutathione S-transferase activity

The Journal of Biological Chemistry
Kun ZhangGuimin Luo

Abstract

Substrate binding and the subsequent reaction are the two principal phenomena that underlie the activity of enzymes, and many enzyme-like catalysts were generated based on the phenomena. The single chain variable region fragment of antibody 2F3 (scFv2F3) was elicited against hapten GSH-S-DN2phBu, a conjugate of glutathione (GSH), butyl alcohol, and 1-chloro-2,4-dinitrobenzene (CDNB); it can therefore bind both GSH and CDNB, the substrates of native glutathione S-transferases (GSTs). It was shown previously that there is a serine residue that is the catalytic group of GST in the CDR regions of scFv2F3 close to the sulfhydryl of GSH. Thus, we anticipated that scFv2F3 will display GST activity. The experimental results showed that scFv2F3 indeed displayed GST activity that is equivalent to the rat-class GST T-2-2 and exhibited pH- and temperature-dependent catalytic activity. Steady-state kinetic studies showed that the Km values for the substrates are close to those of native GSTs, indicating that scFv2F3 has strong affinities for the substrates. Compared with some other GSTs, its kcat value was found to be low, which could be caused by the similarity between the GSH-S-DN2phBu and the reaction product of GSH and CDNB. These resul...Continue Reading

References

Jan 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·A M BensonW B Jakoby
Feb 13, 1990·Biochemistry·R GlockshuberA Plückthun
Jan 1, 1981·Methods in Enzymology·W H Habig, W B Jakoby
Mar 15, 1995·Structure·J L Martin
Feb 15, 1994·Biochemical and Biophysical Research Communications·G M LuoJ C Shen
Jan 1, 1996·Annual Review of Biochemistry·T C Stadtman
Jan 1, 1997·Chemical Research in Toxicology·R N Armstrong
Oct 12, 2000·Nature·J F Atkins, R F Gesteland
May 24, 2001·Archives of Biochemistry and Biophysics·X RenJ Shen
Jul 17, 2001·Biochemical and Biophysical Research Communications·D SuJ Shen
May 9, 2002·Molecular and Cellular Biology·Dolph L Hatfield, Vadim N Gladyshev
May 19, 2004·The Journal of Biological Chemistry·Ye SunGuimin Luo

❮ Previous
Next ❯

Citations

Sep 14, 2011·The International Journal of Biochemistry & Cell Biology·Fei YanGuimin Luo
Sep 23, 2008·The International Journal of Biochemistry & Cell Biology·Yan GeJiacong Shen
Apr 16, 2018·Journal of Molecular Recognition : JMR·Juxin YinYing Mu

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.