Identification of organelles in bacteria similar to acidocalcisomes of unicellular eukaryotes

被引:120
作者
Seufferheld, M
Vieira, MCF
Ruiz, FA
Rodrigues, CO
Moreno, SNJ
Docampo, R
机构
[1] Univ Illinois, Dept Pathobiol, Mol Parasitol Lab, Urbana, IL 61802 USA
[2] Univ Illinois, Ctr Zoonoses Res, Urbana, IL 61802 USA
关键词
D O I
10.1074/jbc.M304548200
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
Acidocalcisomes are acidic calcium storage compartments described in several unicellular eukaryotes, including trypanosomatid and apicomplexan parasites, algae, and slime molds. In this work, we report that the volutin granules of Agrobacterium tumefaciens possess properties similar to the acidocalcisomes. Transmission electron microscopy revealed that each intracellular granule was surrounded by a membrane. X-ray microanalysis of the volutin granules showed large amounts of phosphorus, magnesium, potassium, and calcium. Calcium in the volutin granules increased when the bacteria were incubated at high extracellular calcium concentration. Immunofluorescence and immunoelectron microscopy, using antisera raised against peptide sequences conserved in the A. tumefaciens proton pyrophosphatase, indicated localization in intracellular vacuoles. Purification of the volutin granules using iodixanol density gradients indicated a preferential localization of the pyrophosphatase activity in addition to high concentrations of phosphate, pyrophosphate, short- and long-chain polyphosphate, but lack of markers of the plasma membrane. The pyrophosphatase activity was potassium-insensitive and inhibited by the pyrophosphate analogs, amynomethylenediphosphonate and imidodiphosphate, by dicyclohexylcarbodiimide, and by the thiol reagent N-ethylmaleimide. Polyphosphate was also localized to the volutin granules by 4', 6'-diamino-2-phenylindole staining. The organelles were acidic, as demonstrated by staining with LysoSensor blue DND-167, a dye especially used to detect very acidic compartments in cells, and cycloprodigiosin, a compound isolated from a marine bacterium that has been shown to uncouple proton pyrophosphatase activity acting as a chloride/proton symport. The results suggest that acidocalcisomes arose before the prokaryotic and eukaryotic lineages diverged.
引用
收藏
页码:29971 / 29978
页数:8
相关论文
共 46 条
[1]   INFLUENCE OF S-ADENOSYLMETHIONINE ON DAPI-INDUCED FLUORESCENCE OF POLYPHOSPHATE IN THE YEAST VACUOLE [J].
ALLAN, RA ;
MILLER, JJ .
CANADIAN JOURNAL OF MICROBIOLOGY, 1980, 26 (08) :912-920
[2]   A lysine substitute for K+ -: A460K mutation eliminates K+ dependence in H+-pyrophosphatase of Carboxydothermus hydrogenoformans [J].
Belogurov, GA ;
Lahti, R .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (51) :49651-49654
[3]   H+-Pyrophosphatase of Rhodospirillum rubrum -: High yield expression in Escherichia coli and identification of the Cys residues responsible for inactivation by mersalyl [J].
Belogurov, GA ;
Turkina, MV ;
Penttinen, A ;
Huopalahti, S ;
Baykov, AA ;
Lahti, R .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (25) :22209-22214
[4]   Microcompartments in prokaryotes: Carboxysomes and related polyhedra [J].
Cannon, GC ;
Bradburne, CE ;
Aldrich, HC ;
Baker, SH ;
Heinhorst, S ;
Shively, JM .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2001, 67 (12) :5351-5361
[5]  
COHENBAZIRE G, 1963, BACTERIAL PHOTOSYNTH, P89
[6]   Linearly concatenated cyclobutane lipids form a dense bacterial membrane [J].
Damsté, JSS ;
Strous, M ;
Rijpstra, WIC ;
Hopmans, EC ;
Geenevasen, JAJ ;
van Duin, ACT ;
van Niftrik, LA ;
Jetten, MSM .
NATURE, 2002, 419 (6908) :708-712
[7]   The acidocalcisome [J].
Docampo, R ;
Moreno, SNJ .
MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 2001, 114 (02) :151-159
[8]   A thermostable vacuolar-type membrane pyrophosphatase from the archaeon Pyrobaculum aerophilum:: implications for the origins of pyrophosphate-energized pumps [J].
Drozdowicz, YM ;
Lu, YP ;
Patel, V ;
Fitz-Gibbon, S ;
Miller, JH ;
Rea, PA .
FEBS LETTERS, 1999, 460 (03) :505-512
[9]   Vacuolar H+ pyrophosphatases:: from the evolutionary backwaters into the mainstream [J].
Drozdowicz, YM ;
Rea, PA .
TRENDS IN PLANT SCIENCE, 2001, 6 (05) :206-211
[10]   AVP2, a sequence-divergent, K+-insensitive H+-translocating inorganic pyrophosphatase from arabidopsis [J].
Drozdowicz, YM ;
Kissinger, JC ;
Rea, PA .
PLANT PHYSIOLOGY, 2000, 123 (01) :353-362