Expression and localization of α- and β-carbonic anhydrase in Helicobacter pylori

被引:41
作者
Chirica, LC
Petersson, C
Hurtig, M
Jonsson, BH
Borén, T
Lindskog, S [1 ]
机构
[1] Umea Univ, Dept Chem, SE-90187 Umea, Sweden
[2] Linkoping Univ, Fac Hlth Sci, Div Med Microbiol, Dept Mol & Clin Med, SE-58185 Linkoping, Sweden
[3] Umea Univ, Dept Odontol, SE-90187 Umea, Sweden
[4] Linkoping Univ, Inst Technol, Div Chem, Dept Phys & Measurement Technol, SE-58183 Linkoping, Sweden
来源
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS | 2002年 / 1601卷 / 02期
关键词
expression; localization; carbonic anhydrase;
D O I
10.1016/S1570-9639(02)00467-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Helicobacter pylori, the causative agent of peptic ulcer disease, expresses two different forms of the zinc-containing enzyme carbonic anhydrase (CA) (alpha and beta), catalyzing the reversible hydration Of CO2. Presumably, the high CO2 requirement of H. pylori implies an important role for this enzyme in the bacterial physiology. In this paper, expression of the CAs has been analyzed in three different strains of the bacterium, 26695, J99 and 17.1, and appears to be independent Of CO2 concentration in the investigated range (0.1-10%). Presence of the potent and highly specific CA inhibitor, acetazolamide, in the medium does not seem to inhibit bacterial growth at the given sulfonamide concentration. Moreover, the localization and distribution of the alpha-CA was analyzed by immunonegative staining, while SDS-digested freeze-fracture immunogold labelling was used for the beta-form of the enzyme. The latter method has the advantage of allowing assessment of protein localization to distinct cell compartments and membrane structures. The resulting electron microscopy images indicate a localization of the beta-CA in the cytosol, on the cytosolic side of the inner membrane and on the outer membrane facing the periplasmic space. The alpha-enzyme was found attached to the surface of the bacterium. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:192 / 199
页数:8
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