ANTIBODY GENERATION FOR DETECTION OF PHOSPHOGLYCEROYL PROTEIN ADDUCTS
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Date
2024-04-20
Authors
Shatkenova, Zariat
Journal Title
Journal ISSN
Volume Title
Publisher
Nazarbayev University School of Sciences and Humanities
Abstract
One of the reactions of glycolysis, which is controlled by the enzyme GAPDH, results in the
synthesis of 1,3- bisphosphoglycerate. A highly reactive intermediate product known as
cyclic 1,3-phosphoglycerate (cPGA) can be formed from 1,3-bisphosphoglycerate. In
previous studies, it was shown that the knock-out of PARK7/DJ1 can cause an accumulation
of highly reactive cPGA and its adducts. However, the quantifiable methods to detect cPGA
adducts were lacking. We created a novel technique for the detection of cPGA modifications
with antibodies specifically recognizing cPGA-modified lysine and cysteine residues. We
developed a procedure for the generation of antigens modified by cPGA. We produced
antibodies that recognize lysine and cysteine residues modified by cPGA. Finally, we purified
these antibodies by negative selection with the help of affinity chromatography. Our results
demonstrated that purified antibodies can specifically recognise a diverse range of
cPGA-modified proteins and can be used for Western Blot analysis of cell extracts.
Description
Keywords
cPGA, phospho-glyceroyl protein adducts, DJ 1, GAPDH, Type of access: Restricted
Citation
Shatkenova, Z. (2024).Antibody generation for detection of phosphoglyceroyl protein adducts. Nazarbayev University School of Sciences and Humanities