کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1175953 | 961824 | 2007 | 10 صفحه PDF | دانلود رایگان |

Hypochlorous acid (HOCl) is a major product of activated neutrophils and may be important in antimicrobial activities of cells by oxidation or chlorination of susceptible amino acids. Three major peaks separated using C18 reverse phase-high-performance liquid chromatography RP-HPLC after incubation of leucine enkephalin (LeuEnk) with HOCl. Electrospray mass spectrometry showed masses of m/z 556.2, 590.2, and 624.4 corresponding to unmodified LeuEnk and peptides altered by addition of one or two chlorines (Cl). Formation of stable N-α-chloramines was indicated because the chlorinated peptides were readily reduced with the physiological reductants glutathione and ascorbic acid to LeuEnk (m/z 556.2) within 10 min. Sequence-specific ions observed in product ion spectra of single-charged monochlorinated and dichlorinated peptides were consistent with modification of the N-terminal amine. There was no evidence for chlorination of the Tyr aromatic ring in any spectra. Similar RP-HPLC profiles were obtained after oxidation of des-Tyr1-LeuEnk (GGFL) with the masses of the major products being m/z 393.3, 427.2, and 461.1. These were identified as unmodified GGFL, N-α-Cl-GGFL, and N-α-Cl2-GGFL based on comparison of tandem mass spectra. Oxidation of Met and formation of disulfide dimers was observed after incubation of either N-α-Cl-LeuEnk or N-α-Cl2-LeuEnk with a protein, indicating that both peptide N-α-chloramines were able to readily modify sulfur-containing amino acids within proteins. These data indicate initial formation of stable N-α-chorinated peptides after incubation with HOCl and suggest that N-α-chlorinated peptides may exist for some hours in the absence of physiological reducing agents or sulfur-containing amino acids.
Journal: Analytical Biochemistry - Volume 366, Issue 2, 15 July 2007, Pages 218–227