Probing tau citrullination in Alzheimer’s disease brains and mouse models of tauopathy

Abstract Tauopathies, which include Alzheimer’s disease (AD) share a common defining factor, namely misfolded tau protein. However, the “upstream” etiology and downstream clinical manifestations of tauopathies are quite diverse. Tau deposition elicits different pathological phenotypes and outcomes depending on the tau strain and regional susceptibility. Posttranslational modifications (PTM) can alter tau structure, function, networks, and its pathological sequalae. We uncovered a novel PTM of tau, named citrullination, caused by peptidyl arginine deiminase (PAD) enzymes. PAD induced citrullination irreversibly converts arginine residues to citrulline, producing net loss of positive charge, elimination of pi-pi interactions, and increased hydrophobicity. We observed increased PAD2 and PAD4 in Alzheimer’s disease (AD) brain and that they both can citrullinate tau. Tau can become citrullinated by PADs at all 14 arginine residues throughout the N-terminal domain (N-term), proline-rich domain (PR), microtubule binding repeat domain (MBR), and C-terminal domain (C-term) on full length tau (2N4R). Citrullination of tau impacts fibrillization and oligomerization rates in aggregation assays. Utilizing a panel of novel citrullinated tau (citR tau) antibodies, we identified citrullination of tau in vitro, several animal models of tauopathies, and Alzheimer’s disease (AD). CitR tau increased with Braak stage and was enriched in AD brains with higher phospho-tau burden. This work provides a new area of tau biology that signifies further consideration in the emerging spectrum of tauopathies and its clinical understanding. Competing Interest Statement The authors have declared no competing interest. Footnotes Maj-Linda B. Selenica: Maj-linda.Selenica{at}uky.edu, Jerry B Hunt, Hunt: Jerry.Hunt{at}uky.edu, Huimin Liang: Huimin.Liang{at}uky.edu, Haiying Liu: haiying.liu{at}uky.edu, Junyan Li: junyan.li{at}uky.edu, Malina Serrano: mese235{at}g.uky.edu, Chao Ma: marshall.ma.dmv{at}gmail.com, Andrii Kovalenko: andrii{at}bu.edu, Zainuddin Quadri: Zainuddin.Quadri{at}uky.edu, John Calahatian: johnc1458{at}gmail.com, Cecilie Petersen: cnpedersen{at}mail.usf.edu, Danielle Blazier: dblazier{at}usf.edu, Mallory Watler: mdwatler{at}mail.usf.edu, Patricia Rocha-Rangel: patricia.rocha-rangel{at}uky.edu, Christopher Saunders: christopher.saunders{at}uky.edu, Laura J. Blair: laurablair{at}usf.edu, Leonid Breydo: lbreydo{at}gmail.com, Peter Nelson: pnels2{at}uky.edu, Christopher Norris: christopher.norris{at}uky.edu, Erin Abner: erin.abner{at}uky.edu, Brian Kraemer: kraemerb{at}wu.edu, Kevin Nash: nash{at}usf.edu, Vladimir N. Uversky: vuversky{at}usf.edu, Dale Chaput: chaput{at}usf.edu Abbreviations - PAD - peptidyl arginine deiminase - PTM - post-translational modification - CitR - Citrullination/ citrullinated - AD - Alzheimer’s disease - ALS - Amyotrophic lateral sclerosis - FTD - frontotemporal dementia