Publication:
Proteome and immunome of pathogenic Leptospira spp. revealed by 2DE and 2DE-immunoblotting with immune serum

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Issued Date

2007

Resource Type

Article

File Type

application/pdf

ISSN

0125877X

Other identifier(s)

2-s2.0-34250897884

Rights Holder(s)

Scopus

Bibliographic Citation

Asian Pacific Journal of Allergy and Immunology. Vol 25, No.1 (2007), p.53-73

Suggested Citation

Sakolvaree Y., Maneewatch S., Jiemsup S., Klaysing B., Tongtawe P., Srimanote P., Saengjaruk P., Banyen S., Tapchaisri P., Chonsa-Nguan M., Chaicumpa W. Proteome and immunome of pathogenic Leptospira spp. revealed by 2DE and 2DE-immunoblotting with immune serum. Asian Pacific Journal of Allergy and Immunology. Vol 25, No.1 (2007), p.53-73. Retrieved from: https://hdl.handle.net/20.500.14740/4794

Title

Proteome and immunome of pathogenic Leptospira spp. revealed by 2DE and 2DE-immunoblotting with immune serum

Author(s)

Sakolvaree Y.
 Maneewatch S.
 Jiemsup S.
 Klaysing B.
 Tongtawe P.
 Srimanote P.
 Saengjaruk P.
 Banyen S.
 Tapchaisri P.
 Chonsa-Nguan M.
 Chaicumpa W.

Abstract

In this study, proteomes of two pathogenic Leptospira spp., namely L. interrogans, serogroup Icterohaemorrhagiae, serovar Copenhageni and L. borgpetersenii, serogroup Tarassovi, serovar Tarassovi, were revealed by using two dimensional gel electrophoresis (2DE)-based-proteomics. Bacterial cells were disrupted in a lysis buffer containing 30 mM Tris, 2 M thiourea, 7 M urea, 4% CHAPS, 2% IPG buffer pH 3-10 and protease inhibitors and then subjected to sonication in order to solubilize as much as possible the bacterial proteins. The 2DE-separated components of both Leptospira homogenates were blotted individually onto membranes and antigenic components (immunomes) were revealed by probing the blots with immune serum of a mouse readily immunized with the homogenate of L. interrogans, serogroup Icterohaemorrhagiae, serovar Copenhageni. The immunogenic proteins of the two pathogenic Leptospira spp. could be grouped into 10 groups. These are: 1) proteins involved in the bacterial transcription and translation including beta subunit transcription anti-termination protein of DNA polymerase III, elongation factors Tu and Ts, and tRNA (guanine-N1)-methyltransferase; 2) proteins functioning as enzymes for metabolisms and nutrient acquisition including acetyl-Co-A acetyltransferase, putative glutamine synthetase, glyceraldehyde-3-phospahte dehydrogenase, NifU-like protein, 3-oxoacyl-(acyl-carrier-protein) reductase, oxidoreductase, sphingomyelinase C precursor, spermidine synthase, beta subunit of succinyl-CoA synthetase, and succinate dehydrogenase iron-sulfur subunit; 3) proteins/enzymes necessary for energy and electron transfer, i.e. electron transfer flavoprotein, and proton-translocating transhydrogenase; 4) enzymes for degradation of misfolded proteins, i.e. ATP-dependent Cip protease; 5) molecular chaperone, i.e. 60 kDa chaperonin; 6) signal transduction system, i.e. response regulator; 7) protein involved in immune evasion in host, i.e. peroxiredoxin; 8) cell structure proteins including MreB (cytoskeletal) and flagellin/ periplasmic flagellin; 9) lipoproteins/outer membrane proteins: LipL32, LipL41, LipL45 and OmpL1; and 10) various hypothetical proteins. Many immunogenic proteins are common to both Leptospira spp. These proteins not only are the diagnostic targets but also have potential as candidates of a broad spectrum leptospirosis vaccine especially the surface exposed components which should be vulnerable to the host immune effector factors.

Subject(s)

3 [(3 cholamidopropyl)dimethylammonio] 1 propanesulfonic acid
 3 oxoacyl acyl carrier protein synthase
 Acetyl coenzyme A acetyltransferase
 Bacterial protein
 Bacterial vaccine
 Chaperone
 Chaperonin
 DNA directed DNA polymerase gamma
 Elongation factor Ts
 Elongation factor Tu
 Endopeptidase Clp
 Flavoprotein
 Gene product
 Glutamate ammonia ligase
 Glyceraldehyde 3 phosphate dehydrogenase
 Leptospirosis vaccine
 Lipoprotein
 Nicotinamide adenine dinucleotide (phosphate) transhydrogenase
 NifU like protein
 Outer membrane protein
 Oxidoreductase
 Proteinase inhibitor
 Proteome
 Spermidine synthase
 Sphingomyelin phosphodiesterase
 Succinate dehydrogenase
 Succinyl coenzyme A synthetase
 Transfer RNA methyltransferase
 Unclassified drug
 Urea
 Article
 Bacterial cell
 Blood analysis
 Cell disruption
 Controlled study
 Drug design
 Drug targeting
 Electron transport
 Energy transfer
 Genetic transcription and translation
 Homogenate
 Immune response
 Immunoblotting
 Leptospira
 Leptospira borgpetersenii
 Leptospira interrogans
 Leptospirosis
 Male
 Mouse
 Nonhuman
 Nucleotide sequence
 Protein analysis
 Protein degradation
 Protein folding
 Protein function
 Protein targeting
 Signal transduction
 Species difference
 Two dimensional gel electrophoresis
 Ultrasound
 Amino Acid Sequence
 Animals
 Antigens, Bacterial
 Blotting, Western
 Electrophoresis, Gel, Two-Dimensional
 Leptospira
 Leptospira interrogans serovar icterohaemorrhagiae
 Leptospirosis
 Male
 Mice
 Mice, Inbred BALB C
 Molecular Sequence Data
 Proteome
 Proteomics

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https://hdl.handle.net/20.500.14740/4794

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