Department of Biochemistry and Molecular Pharmacology; Thompson Lab
Amino Acids, Peptides, and Proteins | Biochemical Phenomena, Metabolism, and Nutrition | Biochemistry | Cell Biology | Enzymes and Coenzymes | Molecular Biology | Nucleic Acids, Nucleotides, and Nucleosides | Reproductive and Urinary Physiology
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, alpha-tubulin, and beta-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 microg/mL prolactin for 10 min induced a 2-fold increase in histone H2A citrullination and a 4.5-fold increase in alpha-tubulin citrullination. We next investigated if prolactin-induced citrullination regulates the expression of lactation genes beta-casein (Csn2) and butyrophilin (Btn1a1). Prolactin treatment for 12 h increased beta-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.
citrullination, lactation, mammary gland, peptidylarginine deiminase, prolactin
Rights and Permissions
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
DOI of Published Version
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. Int J Mol Sci. 2020 Apr 10;21(7):2634. doi: 10.3390/ijms21072634. PMID: 32290104; PMCID: PMC7177251. Link to article on publisher's site
International journal of molecular sciences
Li G, Young CH, Snow B, Christensen AO, Demoruelle MK, Nemmara VV, Thompson PR, Rothfuss HM, Cherrington BD. (2020). Identification and Characterization of the Lactating Mouse Mammary Gland Citrullinome. Open Access Publications by UMass Chan Authors. https://doi.org/10.3390/ijms21072634. Retrieved from https://escholarship.umassmed.edu/oapubs/4213
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.
Amino Acids, Peptides, and Proteins Commons, Biochemical Phenomena, Metabolism, and Nutrition Commons, Biochemistry Commons, Cell Biology Commons, Enzymes and Coenzymes Commons, Molecular Biology Commons, Nucleic Acids, Nucleotides, and Nucleosides Commons, Reproductive and Urinary Physiology Commons