Cell cycle arrest by monochloramine through the oxidation of retinoblastoma protein. 
Impairment of cell cycle control has serious effects on inflammation, tissue repair, and carcinogenesis.  We report here the G1 cell cycle arrest by monochloramine (NH2Cl), a physiological oxidant derived from activated neutrophils, and its mechanism.  When Jurkat cells were treated with NH2Cl (70 microM, 10 min) and incubated for 24 h, the S phase population decreased significantly with a slight increase in the hypodiploid cell population.  The G0/ G1 phase and G2/M phase populations did not show marked changes.  Three hours after NH2Cl treatment, the retinoblastoma protein (pRB) was dephosphorylated especially at Ser780 and Ser795, both of which are important phosphorylation sites for the G1 checkpoint function.  The phosphorylation at Ser807/811 showed no apparent change.  The expression of cyclins, cyclin-dependent kinases, and cyclin-dependent kinase inhibitors showed no apparent change.  Moreover, the kinase activity that phosphorylates pRB remained constant even after NH2Cl treatment.  The protein phosphatase activity that dephosphorylates pRB showed a marginal increase.  Notably, when the recombinant pRB was oxidized by NH2Cl in vitro, the oxidized pRB became difficult to be phosphorylated by kinases, especially at Ser780 and Ser795, but not at Ser807/811.  Amino acid analysis of oxidized pRB showed methionine oxidation to methionine sulfoxide.  The NH2Cl-treated Jurkat cell proteins also showed a decrease in methionine.  These observations suggested that direct pRB oxidation was the major cause of NH2Cl-induced cell cycle arrest.  In the presence of 2 mM NH4+, NaOCl (200 microM) or activated neutrophils also induced a G1 cell cycle arrest.  As protein methionine oxidation has been reported in inflammation and aging, cell cycle modulation by pRB oxidation may occur in various pathological conditions.