ADP-RIBOSYLTRANSFERASE OF HELIOBACTER PYLORI

幽门螺杆菌 ADP-核糖基转移酶

• 批准号: 09044322
• 负责人: HIRAYAMA Toshiya
• 金额: $ 4.99万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09044322/
• 关键词: HELICOBACTER PYLORI; ADP-RIBOSYLTRANSFERASE; BACTERIAL TOXIN; ETIOLOGIC AGENT; BACTERIAL INFECTION

HELICOBACTER PYLORI IS AN ETIOLOGIC AGENT FOR CHRONlC GASTRITIS. DUODENAL ULCERS. AND GASTRIC MUCOUS MALIGNANCY. TO VESTIGATE FACTORS INVOLVED IN THE PATHOGENESIS OF THE MICROORGANISM. WE EXAMlNED WHETHER H. PYLORI PRODUCES AN ADP-RIBOSYLTRANSFERASE (ART) AND WHETHER AN ANTI-ULCER DRUG CAN INHIBIT THE ENZYME ACTIVITY. SONIFIED CULTURE OF THE BACTERIA WAS SUBJECTED TO ART ASSAY IN WHICH THE EXTRACT WAS INCUBATED WITH [ADENYLATE-32P]NAD. WHEN THE ASSAY MIXTURE WAS ANALYZED BY SDS-PAGE AND AUTORADIOGRAPHY, RADIOLABELELLING OF A 70KDA PROTEIN (P70) IN THE CELL LYSATE WAS OBSERVED IN A MG2+-DEPENDENT MANNER. THE MODIFICATION OF P70 SEEMED TO BE MEDIATED BY A MONO-ART, AND INVOLVEMENT OF NON-ENZYMATIC NAD-DEPENDENT MODIFICATIONS APPEARED TO BE UNLIKELY. THE OBSERVATIONS THAT (ADP-RIBOSE)-P70 LINKAGE WAS SUSCEPTIBLE TO HYDROXYLAMlNE TREATMENT AND THAT ARGlNlNE AND AGMATINE INHIBITED THE MODIFICATION OF P70 INDICATED THAT ARGlNlNE RESIDUE IN P70 IS THE SITE OF THE MODIFICATION. ADDITIONALLY, AGMATINE COULD BE UTILIZED AS A SUBSTRATE BY THE PUTATIVE ENZYME IN A MG2+-DEPENDENT MANNER. H. PYLORI ART WAS NOT ACTIVATED BY ARF, A SMALL GTP-BINDlNG PROTEIN WHICH IS STIMULATE ART ACTIVITY OF CHOLERA ENTEROTOXlN. AN ANTI-ULCER COMPOUND, REBAMIPIDE, INHIBITED THE ADP-RIBOSYLATION BOTH OF P70 AND OF AGMATINE BY H. PYLORI CELL LYSATE. SUGGESTlNG THAT THE ENZYME CAN BE A TARGET OF THE DRUG. THESE RESULTS INDICATE THAT H. PYLORI PRODUCES A CELL-ASSOCIATED ARGlNlNE-SPECIFIC MONO-ART, WHICH IS SUSCEPTIBLE TO ANTI-ULCER DRUG. FURTHERMORE. INCUBATION OF H. PYLORI LYSATE WITH THE MOMOGENATE OF HL-60 CELLS CAUSED THE MODIFICATION OF A 50KDA PROTElN, SUGGESTING THAT THE ENZYME MAY ALSO ADP-RIBOSYLATE HOST CELL PROTEIN. BECAUSE MANY BACTERIAL PATHOGEN PRODUCE TOXINS WITH MONO-ART ACTIVITY, THE RESULTS MAY SUGGEST THAT H. PYLORI ART PLAYS A ROLE IN PATHOLOGICAL EFFECT OF THE BACTERlUM.

幽门螺杆菌是慢性胃炎的病原体十二指肠溃疡。和胃粘膜恶性肿瘤。探讨微生物致病因素。我们怀疑H。幽门螺杆菌产生ADP-核糖基转移酶（ART）以及抗溃疡药物是否能抑制该酶活性。对芽孢杆菌的超声培养物进行ART试验，其中浸提液与[腺苷酸-32 P]NAD孵育。当通过SDS-PAGE和放射自显影法分析试验混合物时，以MG 2+依赖性方式观察到细胞裂解物中70 kDa蛋白（P70）的放射性标记。P70的修饰似乎是由一个单一的艺术介导的，涉及非酶NAD依赖性修饰似乎是不可能的。观察到（ADP-核糖）-P70连接对羟胺处理敏感，精氨酸和胍基丁胺抑制P70的修饰，表明P70中的精氨酸残基是修饰的位点。此外，胍丁胺可以以MG 2+依赖性方式被假定的酶用作底物。H.幽门螺旋杆菌ART不被ARF激活，ARF是一种刺激霍乱毒素ART活性的小GTP结合蛋白。抗溃疡化合物瑞巴派特抑制H.幽门细胞裂解物。表明酶可能是药物的靶点。这些结果表明，H。幽门螺杆菌产生细胞相关的精氨酸特异性单抗，其对抗溃疡药物敏感。更进一步。H.用HL-60细胞的原核细胞裂解幽门螺杆菌，引起50 kDa蛋白质的修饰，表明该酶也能使宿主细胞蛋白ADP-核糖基化。由于许多细菌病原体产生的毒素具有单一的活性，结果可能表明，H。幽门螺杆菌艺术发挥作用的病理效应的钡。

项目成果

YASHIRO,Kinnosuke, NIIDOME,Takuro, HATAKEYAMA,Tomomitsu, AOYAGI,Haruhiko, KURAZONO,Hisao, Philip Ian Padilla, WADA,Akihiro, HIRAYAMA,Toshiya: "Helicobacter pylori vacuolating cytotoxin (VacA) binds to the 140kDa protein in human gastric cancer cell lines,

YASHIRO、Kinnosuke、NIIDOME、Takuro、HATAKEYAMA、Tomomitsu、AOYAGI、Haruhiko、KURAZONO、Hisao、Philip Ian Padilla、WADA、Akihiro、HIRAYAMA、Toshiya：“幽门螺杆菌空泡细胞毒素 (VacA) 与人胃癌细胞中的 140kDa 蛋白结合

Naoya Ohmori et al.: "Importance of hydrophobic region in amphiphilic structures of α-helical peptides for their gene transfer-ability into cells." Biochem.Biophys.Res.Commun.245. 259-265 (1998)

Naoya Ohmori 等人：“α-螺旋肽的两亲结构中的疏水区域对于其基因转移到细胞中的能力的重要性。”Biochem.Biophys.Res.Commun.245 (1998)。

KIMURA,Miyuki, GOTO,Shinji, WADA,Akihiro, YAHIRO,Kinosuke, NIIDOME,Takuro, HATAKEYAMA,Tomomitsu, AOYAGI,Haruhiko, HIRAYAMA,Toshiya, and KONDO,Takahito: "Vacuolating cytotoxin purified from Helicobacter pylori causes mitochondorial damage in human gastric

KIMURA、Miyuki、GOTO、Shinji、WADA、Akihiro、YAHIRO、Kinosuke、NIIDOME、Takuro、HATAKEYAMA、Tomomitsu、AOYAGI、Haruhiko、HIRAYAMA、Toshiya 和 KONDO、Takahito：“从幽门螺杆菌中纯化的空泡细胞毒素会导致人胃线粒体损伤

Oishi, K.et al.: "Nitrite reductase from Pseudomonas aeruginosa induces inflammatory cvtokines in cultured respiratory cells." Infect . Immun .65. 2648-2655 (1997)

Oishi, K. 等人：“来自铜绿假单胞菌的亚硝酸盐还原酶在培养的呼吸细胞中诱导炎症细胞因子。”

HIRAYAMA, Toshiya: "Functions of pathogenic factors from Helicobacter pylori"Helicobacter Research. 4. 379-379 (1997)

HIRAYAMA、Toshiya：“幽门螺杆菌致病因子的功能”螺杆菌研究。