Citrullination of adenosine deaminase impairs its binding to dipeptidyl peptidase IV

Luiza Karapetyan; Svetlana Sharoyan; Sona Mardanyan; Giulio Lupidi; Massimiliano Cuccioloni; Mauro Angeletti; Shiraz Markarian; Hasmik Shilajyan; Alvard Antonyan

doi:10.1016/j.bpc.2022.106820

Citrullination of adenosine deaminase impairs its binding to dipeptidyl peptidase IV

Biophys Chem. 2022 Jul:286:106820. doi: 10.1016/j.bpc.2022.106820. Epub 2022 Apr 15.

Authors

Luiza Karapetyan¹, Svetlana Sharoyan², Sona Mardanyan², Giulio Lupidi³, Massimiliano Cuccioloni⁴, Mauro Angeletti⁵, Shiraz Markarian⁶, Hasmik Shilajyan⁷, Alvard Antonyan¹

Affiliations

¹ H. Buniatian Institute of Biochemistry of Armenian, NAS, 5/1, P. Sevak Str., Yerevan 0014, Armenia. Electronic address: biochem@biochem.sci.am.
² H. Buniatian Institute of Biochemistry of Armenian, NAS, 5/1, P. Sevak Str., Yerevan 0014, Armenia.
³ School of Pharmacy, University of Camerino, via S. Agostino 1, 62032 Camerino, MC, Italy. Electronic address: giulio.lupidi@unicam.it.
⁴ School of Biosciences and Veterinary Medicine, Via Gentile III da Varano, 62032 Camerino, MC, Italy. Electronic address: massimiliano.cuccioloni@unicam.it.
⁵ School of Biosciences and Veterinary Medicine, Via Gentile III da Varano, 62032 Camerino, MC, Italy.
⁶ Faculty of Chemistry, Yerevan State University, Yerevan, Armenia. Electronic address: shmarkar@ysu.am.
⁷ Faculty of Chemistry, Yerevan State University, Yerevan, Armenia.

PMID: 35468399
DOI: 10.1016/j.bpc.2022.106820

Abstract

The presence of citrullinated adenosine deaminase (ADA) was reported in the synovial fluids of rheumatoid arthritis individuals. This work reports the effects of ADA citrullination on the formation/stabilization of ADA complex with dipeptidyl peptidase IV (DPPIV). The electrophoretic mobility of in vivo citrullinated ADA was diminished compared to the native one. The biosensor binding study demonstrated approximately four-fold lower affinity of both in vivo and in vitro citrullinated ADAs to DPPIV (K_D = 161 ± 51.3 and 171 ± 52.2 nM, respectively) compared with wild ADA (K_D = 38 ± 9.4 nM). These results were confirmed by examining the ADA interaction with DPPIV using size-exclusion chromatography and fluorescence anisotropy methods. The computational modeling of Arg142 → Cit142 modification in ADA showed a local structural rearrangement and a less favorable binding affinity to DPPIV. According to these observations, citrullinated ADA being a possible target triggering autoimmunity hinders also the formation of ADA-DPPIV complex, essential in immune system function.

Keywords: Citrullinated adenosine deaminase; Computational modeling; Electrophoretic mobility; Fluorescence anisotropy; Protein-protein interaction; Resonant mirror biosensor assay.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenosine Deaminase* / chemistry
Adenosine Deaminase* / metabolism
Citrullination*
Dipeptidyl Peptidase 4* / chemistry
Dipeptidyl Peptidase 4* / genetics
Dipeptidyl Peptidase 4* / metabolism
Humans

Substances

DPP4 protein, human
Dipeptidyl Peptidase 4
ADA protein, human
Adenosine Deaminase