中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2014年
2期
91-95
,共5页
潘建义%王长一%叶智鸧%陈冉%赵辅昆
潘建義%王長一%葉智鸧%陳冉%趙輔昆
반건의%왕장일%협지창%진염%조보곤
铜绿假单胞菌%巨噬细胞%应激%蛋白质磷酸化%磷酸化蛋白质组%致病机制
銅綠假單胞菌%巨噬細胞%應激%蛋白質燐痠化%燐痠化蛋白質組%緻病機製
동록가단포균%거서세포%응격%단백질린산화%린산화단백질조%치병궤제
Pseudomonas aeruginosa%Macrophage%Bacterial stress response%Protein phosphorylation%Phosphoproteome%Pathongenesis
目的 鉴定巨噬细胞应激的铜绿假单胞菌(Pseudomonas aeruginosa)磷酸化蛋白质组,分析磷酸化修饰在该菌应答应激中的作用.方法 采用强阳离子交换层析和固相金属离子亲和层析(SCX-IMAC)富集应激细菌的磷酸化肽段,并利用纳升级液相色谱-质谱联用技术(nano LC-MS/MS)进行鉴定和分析其磷酸化蛋白质组.结果 在应激细菌中鉴定到14个磷酸化肽段,对应于12个磷酸化蛋白质,其中膜蛋白占50％,提示膜蛋白的磷酸化修饰在这一过程中具有关键作用.蛋白质功能分析显示这些磷酸化蛋白质主要涉及应激应答、铁摄取转运、厌氧呼吸、过氧化氢的应激应答、磷酸化介导的信号转导等生物过程.结论 铜绿假单胞菌的蛋白质磷酸化修饰对其转导应激信号及应答巨噬细胞内的缺铁、缺氧和氧化应激等不利生存环境具有关键作用,该结果为深入理解该菌的毒力及致病机制提供新的思路.
目的 鑒定巨噬細胞應激的銅綠假單胞菌(Pseudomonas aeruginosa)燐痠化蛋白質組,分析燐痠化脩飾在該菌應答應激中的作用.方法 採用彊暘離子交換層析和固相金屬離子親和層析(SCX-IMAC)富集應激細菌的燐痠化肽段,併利用納升級液相色譜-質譜聯用技術(nano LC-MS/MS)進行鑒定和分析其燐痠化蛋白質組.結果 在應激細菌中鑒定到14箇燐痠化肽段,對應于12箇燐痠化蛋白質,其中膜蛋白佔50％,提示膜蛋白的燐痠化脩飾在這一過程中具有關鍵作用.蛋白質功能分析顯示這些燐痠化蛋白質主要涉及應激應答、鐵攝取轉運、厭氧呼吸、過氧化氫的應激應答、燐痠化介導的信號轉導等生物過程.結論 銅綠假單胞菌的蛋白質燐痠化脩飾對其轉導應激信號及應答巨噬細胞內的缺鐵、缺氧和氧化應激等不利生存環境具有關鍵作用,該結果為深入理解該菌的毒力及緻病機製提供新的思路.
목적 감정거서세포응격적동록가단포균(Pseudomonas aeruginosa)린산화단백질조,분석린산화수식재해균응답응격중적작용.방법 채용강양리자교환층석화고상금속리자친화층석(SCX-IMAC)부집응격세균적린산화태단,병이용납승급액상색보-질보련용기술(nano LC-MS/MS)진행감정화분석기린산화단백질조.결과 재응격세균중감정도14개린산화태단,대응우12개린산화단백질,기중막단백점50％,제시막단백적린산화수식재저일과정중구유관건작용.단백질공능분석현시저사린산화단백질주요섭급응격응답、철섭취전운、염양호흡、과양화경적응격응답、린산화개도적신호전도등생물과정.결론 동록가단포균적단백질린산화수식대기전도응격신호급응답거서세포내적결철、결양화양화응격등불리생존배경구유관건작용,해결과위심입리해해균적독력급치병궤제제공신적사로.
Objective To investigate the role of protein phosphorylation in Pseudomonas aeruginosa (P.aeruginosa) strains in response to stress triggered by mouse macrophages.Methods The strong cation exchange-immobilized metal affinity chromatography (SCX-IMAC) was performed to enrich phosphopeptides.The nanoscale liquid chromatography coupled to tandem mass spectrometry (nano LC-MS/MS) was carried out to identify and analyze phosphoproteome.Results Fourteen phosphopeptides from twelve proteins were identified within thirty-one phosphorylation sites on serine,threonine and tyrosine residues.Fifty percent of these phosphorylated proteins were membrane proteins,indicating that their phosphorylation modification was more critical for bacteria in response to the stress.In terms of biological process of Gene Ontology,these identified proteins were involved in stress response,iron transport,anaerobic respiration,response to hydrogen peroxide and signal transduction by phosphorylation,etc.Conclusion These phosphorylated proteins in P.aeruginosa strains are necessary for signal transduction and their response to harsh environment within the macrophages,such as iron limitation,hypoxia and oxidative stress.This study provides evidence for further investigation on virulence and pathogenesis of P.aeruginosa.
