Chemo-enzymatic detection of protein isoaspartate using protein isoaspartate methyltransferase and hydrazine trapping.

Analytical Chemistry
Joshua F AlfaroZhaohui Sunny Zhou

Abstract

Isoaspartate formation is a ubiquitous post-translation modification arising from spontaneous asparagine deamidation or aspartate isomerization. The formation of isoaspartate inserts a methylene group into the protein backbone, generating a "kink", and may drastically alter protein structure and function, thereby playing critical roles in a myriad of biological processes, human diseases, and protein pharmaceutical development. Herein, we report a chemo-enzymatic detection method for the isoaspartate protein, which in particular allows the affinity enrichment of isoaspartate-containing proteins. In the initial step, protein isoaspartate methyltransferase selectively converts isoaspartates into the corresponding methyl esters. Subsequently, the labile methyl ester is trapped by strong nucleophiles in aqueous solutions, such as hydrazines to form hydrazides. The stable hydrazide products can be analyzed by standard proteomic techniques, such as matrix-assisted laser desorption ionization and electrospray ionization mass spectrometry. Furthermore, the chemical trapping step allows us to introduce several tagging strategies for product identification and quantification, such as UV-vis and fluorescence detection through a dansyl deri...Continue Reading

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Citations

Jul 9, 2011·Analytical Chemistry·Nadezda P SargaevaPeter B O'Connor
Jul 22, 2014·Journal of Pharmaceutical Sciences·Joshua J KlaeneZhaohui Sunny Zhou
Jan 15, 2016·Mass Spectrometry Reviews·Piliang HaoSiu Kwan Sze
Dec 9, 2015·International Journal of Mass Spectrometry·Xiang YuCheng Lin
May 26, 2015·Journal of Sulphur Chemistry·Richard I DuclosZhaohui Sunny Zhou
Aug 5, 2016·Journal of the American Society for Mass Spectrometry·Pradeep G KabadiAmarnath Chatterjee
Sep 13, 2016·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Tianzhu ZangJisheng Ma
Feb 22, 2017·Nature Chemistry·Megan L MatthewsBenjamin F Cravatt
Jun 27, 2019·Biochemistry. Biokhimii︠a︡·P K K Mishra, M Mahawar
Mar 2, 2021·Antibody Therapeutics·Amissi SadikiZhaohui Sunny Zhou
Mar 17, 2018·Journal of the American Society for Mass Spectrometry·S T AyrtonR G Cooks
Feb 2, 2012·Bioconjugate Chemistry·Jie LiuYing Luo
Aug 20, 2014·Analytical Chemistry·Chris ChumsaeZhaohui Sunny Zhou

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