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Article

Identification and Characterization of the Lactating Mouse Mammary Gland Citrullinome

1
Department of Zoology and Physiology, University of Wyoming, Laramie, WY 82071, USA
2
Division of Rheumatology, University of Colorado School of Medicine, Aurora, CO 80045, USA
3
Department of Chemistry and Biochemistry, Rowan University, Glassboro, NJ 08028, USA
4
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605, USA
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(7), 2634; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21072634
Received: 12 March 2020 / Revised: 7 April 2020 / Accepted: 8 April 2020 / Published: 10 April 2020
Citrullination is a post-translational modification (PTM) in which positively charged peptidyl-arginine is converted into neutral peptidyl-citrulline by peptidylarginine deiminase (PAD or PADI) enzymes. The full protein citrullinome in many tissues is unknown. Herein, we used mass spectrometry and identified 107 citrullinated proteins in the lactation day 9 (L9) mouse mammary gland including histone H2A, α-tubulin, and β-casein. Given the importance of prolactin to lactation, we next tested if it stimulates PAD-catalyzed citrullination using mouse mammary epithelial CID-9 cells. Stimulation of CID-9 cells with 5 µg/mL prolactin for 10 min induced a 2-fold increase in histone H2A citrullination and a 4.5-fold increase in α-tubulin citrullination. We next investigated if prolactin-induced citrullination regulates the expression of lactation genes β-casein (Csn2) and butyrophilin (Btn1a1). Prolactin treatment for 12 h increased β-casein and butyrophilin mRNA expression; however, this increase was significantly inhibited by the pan-PAD inhibitor, BB-Cl-amidine (BB-ClA). We also examined the effect of tubulin citrullination on the overall polymerization rate of microtubules. Our results show that citrullinated tubulin had a higher maximum overall polymerization rate. Our work suggests that protein citrullination is an important PTM that regulates gene expression and microtubule dynamics in mammary epithelial cells. View Full-Text
Keywords: peptidylarginine deiminase; citrullination; prolactin; mammary gland; lactation peptidylarginine deiminase; citrullination; prolactin; mammary gland; lactation
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MDPI and ACS Style

Li, G.; Young, C.H.; Snow, B.; Christensen, A.O.; Demoruelle, M.K.; Nemmara, V.V.; Thompson, P.R.; Rothfuss, H.M.; Cherrington, B.D. Identification and Characterization of the Lactating Mouse Mammary Gland Citrullinome. Int. J. Mol. Sci. 2020, 21, 2634. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21072634

AMA Style

Li G, Young CH, Snow B, Christensen AO, Demoruelle MK, Nemmara VV, Thompson PR, Rothfuss HM, Cherrington BD. Identification and Characterization of the Lactating Mouse Mammary Gland Citrullinome. International Journal of Molecular Sciences. 2020; 21(7):2634. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21072634

Chicago/Turabian Style

Li, Guangyuan, Coleman H. Young, Bryce Snow, Amanda O. Christensen, M. K. Demoruelle, Venkatesh V. Nemmara, Paul R. Thompson, Heather M. Rothfuss, and Brian D. Cherrington 2020. "Identification and Characterization of the Lactating Mouse Mammary Gland Citrullinome" International Journal of Molecular Sciences 21, no. 7: 2634. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21072634

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