Department of Respiratory, Inflammation, and Autoimmunity, MedImmune LLC, Gaithersburg, MD, United States.
Front Immunol. 2018 Jan 18;9:34. doi: 10.3389/fimmu.2018.00034. eCollection 2018.
Protein citrullination catalyzed by peptidyl arginine deiminase (PADs) is involved in autoimmune disease pathogenesis, especially in rheumatoid arthritis. Calcium is a key regulator of PAD activity, but under normal physiological conditions it remains uncertain how intracellular calcium levels can be raised to sufficiently high levels to activate these enzymes. In pursuit of trying to identify other factors that influence PAD activity, we identified bicarbonate as a potential regulator of PAD activity. We demonstrate that physiological levels of bicarbonate upregulate citrullination by recombinant PAD2/4 and endogenous PADs in neutrophils. The impact of bicarbonate is independent of calcium and pH. Adding bicarbonate to commercial PAD activity kits could increase assay performance and biological relevance. These results suggest that citrullination activity is regulated by multiple factors including calcium and bicarbonate. We also provide commentary on the current understanding of PAD regulation and future perspective of research in this area.
肽基精氨酸脱亚氨酶(PADs)催化的蛋白质瓜氨酸化参与自身免疫性疾病的发病机制,尤其是类风湿关节炎。钙是 PAD 活性的关键调节剂,但在正常生理条件下,尚不清楚如何将细胞内钙水平提高到足以激活这些酶的水平。在试图确定其他影响 PAD 活性的因素时,我们发现碳酸氢盐是 PAD 活性的潜在调节剂。我们证明生理浓度的碳酸氢盐可上调中性粒细胞中重组 PAD2/4 和内源性 PADs 的瓜氨酸化。碳酸氢盐的作用独立于钙和 pH 值。向商业 PAD 活性试剂盒中添加碳酸氢盐可以提高测定性能和生物学相关性。这些结果表明,瓜氨酸化活性受到多种因素的调节,包括钙和碳酸氢盐。我们还对 PAD 调节的现有理解以及该领域未来的研究方向进行了评论。