Triton X‐114 Fractionated Subcellular Proteome of Leptospira interrogans Shows Selective Enrichment of Pathogenic and Outer Membrane Proteins in the Detergent Fraction

PROTEOMICS ◽  
2020 ◽  
Vol 20 (19-20) ◽  
pp. 2000170
Author(s):  
Sikha Thoduvayil ◽  
Gunasekaran Dhandapani ◽  
Rahul Brahma ◽  
Rex Devasahayam Arokia Balaya ◽  
Kiran K. Mangalaparthi ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Outer Membrane ◽  
Outer Membrane Proteins ◽  
Leptospira Interrogans ◽  
Selective Enrichment ◽  
Triton X

scholarly journals Correction: Fine Mapping of the Interaction between C4b-Binding Protein and Outer Membrane Proteins LigA and LigB of Pathogenic Leptospira interrogans

2015 ◽  
Vol 9 (12) ◽  
pp. e0004286 ◽  
Author(s):  
Leandro C. D. Breda ◽  
Ching-Lin Hsieh ◽  
Mónica M. Castiblanco Valencia ◽  
Ludmila B. da Silva ◽  
Angela S. Barbosa ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Outer Membrane ◽  
Fine Mapping ◽  
Binding Protein ◽  
Outer Membrane Proteins ◽  
Leptospira Interrogans ◽  
Pathogenic Leptospira

scholarly journals Fine Mapping of the Interaction between C4b-Binding Protein and Outer Membrane Proteins LigA and LigB of Pathogenic Leptospira interrogans

2015 ◽  
Vol 9 (10) ◽  
pp. e0004192 ◽  
Author(s):  
Leandro C. D. Breda ◽  
Ching-Lin Hsieh ◽  
Mónica M. Castiblanco Valencia ◽  
Ludmila B. da Silva ◽  
Angela S. Barbosa ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Outer Membrane ◽  
Fine Mapping ◽  
Binding Protein ◽  
Outer Membrane Proteins ◽  
Leptospira Interrogans ◽  
Pathogenic Leptospira

Molecular characterization of the pL40 protein inLeptospira interrogans

2009 ◽  
Vol 55 (6) ◽  
pp. 739-749
Author(s):  
Wei Zhao ◽  
Chun-Yan Chen ◽  
Xiang-Yan Zhang ◽  
Wei-Qiang Lai ◽  
Bao-Yu Hu ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Vaccine Candidate ◽  
Outer Membrane Proteins ◽  
Leptospira Interrogans ◽  
Major Focus ◽  
Pcr Screening ◽  
Low Passage ◽  
Mammalian Infection ◽  
Triton X

Leptospirosis is a widespread zoonotic disease caused by pathogenic leptospires. The identification of outer membrane proteins (OMPs) conserved among pathogenic leptospires, which are exposed on the leptospiral surface and expressed during mammalian infection, has become a major focus of leptospirosis research. pL40, a 40 kDa protein coded by the LA3744 gene in Leptospira interrogans , was found to be unique to Leptospira . Triton X-114 fractionation and flow cytometry analyses indicate that pL40 is a component of the leptospiral outer membrane. The conservation of pL40 among Leptospira strains prevalent in China was confirmed by both Western blotting and PCR screening. Furthermore, the pL40 antigen could be recognized by sera from guinea pigs and mice infected with low-passage L. interrogans. These findings indicate that pL40 may serve as a useful serodiagnostic antigen and vaccine candidate for L. interrogans.

scholarly journals Temperature-Regulated Protein Synthesis by Leptospira interrogans

2001 ◽  
Vol 69 (1) ◽  
pp. 400-404 ◽  
Author(s):  
Jarlath E. Nally ◽  
John F. Timoney ◽  
Brian Stevenson
Keyword(s):  
Protein Synthesis ◽  
Outer Membrane ◽  
Pathogenic Bacteria ◽  
Outer Membrane Proteins ◽  
Pathogen Transmission ◽  
Leptospira Interrogans ◽  
Outer Membrane Lipoprotein ◽  
Higher Temperature ◽  
Response To Temperature ◽  
Triton X

ABSTRACT Leptospira interrogans is an important mammalian pathogen. Transmission from an environmental source requires adaptations to a range of new environmental conditions in the organs and tissues of the infected host. Since many pathogenic bacteria utilize temperature to discern their environment and regulate the synthesis of appropriate proteins, we investigated the effects of temperature on protein synthesis in L. interrogans. Bacteria were grown for several days after culture temperatures were shifted from 30 to 37°C. Triton X-114 cellular fractionation identified several proteins of the cytoplasm, periplasm, and outer membrane for which synthesis was dependent on the culture temperature. Synthesis of a cytoplasmic protein of 20 kDa was switched off at 37°C, whereas synthesis of a 66-kDa periplasmic protein was increased at the higher temperature. Increased synthesis of a 25-kDa outer membrane protein was observed when the organisms were shifted from 30 to 37°C. A 36-kDa protein synthesized at 30 but not at 37°C was identified as LipL36, an outer membrane lipoprotein. In contrast, expression of another lipoprotein, LipL41, was the same at either temperature. Immunoblotting with convalescent equine sera revealed that some proteins exhibiting thermoregulation of synthesis elicited antibody responses during infection. Our results show that sera from horses which aborted as a result of naturally acquired infection withL. interrogans serovar pomona type kennewicki recognize periplasmic and outer membrane proteins which are differentially synthesized in response to temperature and which therefore may be important in the host-pathogen interaction during infection.

scholarly journals Protein and antigen profiles of Leptospira interrogans serovar Hardjo

2009 ◽  
Vol 39 (9) ◽  
pp. 2539-2543
Author(s):  
Bárbara Nobre Lafetá ◽  
Elaine Cristina de Castro ◽  
Nivaldo da Silva
Keyword(s):  
Immune Response ◽  
Membrane Proteins ◽  
Western Blot ◽  
Outer Membrane ◽  
Protein Profile ◽  
Outer Membrane Proteins ◽  
Rabbit Antiserum ◽  
Leptospira Interrogans ◽  
Sds Page ◽  
Hyperimmune Rabbit

The protein profile of the outer membrane of Leptospira interrogans serovar Hardjo subtype hardjoprajitno associated with the bovine natural immune response was investigated. The outer membrane proteins were extracted utilizing Triton X114 and precipitated with acetone. The protein sample was then resolved by SDS-PAGE and reacted in western blot against sera from a hyperimmune rabbit and from naturally infected bovines. In silver stained gels, 14 protein bands were observed, among which four proteins, with 22, 29, 47 and 63kDa, appeared as major constituents. Western blot tests with hyperimmune rabbit antiserum detected bands corresponding to proteins with 35; 27; 24; 21; 17 and 14kDa, while 32kDa and 45kDa proteins were the most immunoreactive with sera from naturally infected bovines.

scholarly journals Protective Immunity and Reduced Renal Colonization Induced by Vaccines Containing Recombinant Leptospira interrogans Outer Membrane Proteins and Flagellin Adjuvant

2015 ◽  
Vol 22 (8) ◽  
pp. 965-973 ◽  
Author(s):  
D. Monaris ◽  
M. E. Sbrogio-Almeida ◽  
C. C. Dib ◽  
T. A. Canhamero ◽  
G. O. Souza ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Immune Responses ◽  
Outer Membrane ◽  
Protective Immunity ◽  
Outer Membrane Proteins ◽  
Protein A ◽  
Leptospira Interrogans ◽  
Renal Tubules ◽  
Subunit Vaccines ◽  
Content Type

ABSTRACTLeptospirosis is a global zoonotic disease caused by differentLeptospiraspecies, such asLeptospira interrogans, that colonize the renal tubules of wild and domestic animals. Thus far, attempts to develop effective leptospirosis vaccines, both for humans and animals, have failed to induce immune responses capable of conferring protection and simultaneously preventing renal colonization. In this study, we evaluated the protective immunity induced by subunit vaccines containing seven different recombinantLeptospira interrogansouter membrane proteins, including the carboxy-terminal portion of the immunoglobulinlike protein A (LigAC) and six novel antigens, combined with aluminum hydroxide (alum) orSalmonellaflagellin (FliC) as adjuvants. Hamsters vaccinated with the different formulations elicited high antigen-specific antibody titers. Immunization with LigAC, either with alum or flagellin, conferred protective immunity but did not prevent renal colonization. Similarly, animals immunized with LigACor LigACcoadministered with six leptospiral proteins with alum adjuvant conferred protection but did not reduce renal colonization. In contrast, immunizing animals with the pool of seven antigens in combination with flagellin conferred protection and significantly reduced renal colonization by the pathogen. The present study emphasizes the relevance of antigen composition and added adjuvant in the efficacy of antileptospirosis subunit vaccines and shows the complex relationship between immune responses and renal colonization by the pathogen.

scholarly journals Outer Membrane Proteins Omp10, Omp16, and Omp19 of Brucella spp. Are Lipoproteins

1999 ◽  
Vol 67 (9) ◽  
pp. 4960-4962 ◽  
Author(s):  
Anne Tibor ◽  
Béatrice Decelle ◽  
Jean-Jacques Letesson
Keyword(s):  
Monoclonal Antibodies ◽  
Membrane Proteins ◽  
Palmitic Acid ◽  
Outer Membrane ◽  
Brucella Abortus ◽  
Outer Membrane Proteins ◽  
Specific Inhibitor ◽  
Signal Peptidase ◽  
Processing Sequence ◽  
Triton X

ABSTRACT The deduced sequences of the Omp10, Omp16, and Omp19 outer membrane proteins of Brucella spp. contain a potential bacterial lipoprotein processing sequence. After extraction with Triton X-114, these three proteins partitioned into the detergent phase. Processing of the three proteins is inhibited by globomycin, a specific inhibitor of lipoprotein signal peptidase. The three proteins were radioimmunoprecipitated from [3H]palmitic acid-labeledBrucella abortus lysates with monoclonal antibodies. These results demonstrate that Omp10, Omp16, and Omp19 are lipoproteins.

scholarly journals Expression and Comparative Analysis of Genes Encoding Outer Membrane Proteins LipL21, LipL32 and OmpL1 in Epidemic Leptospires

2005 ◽  
Vol 37 (10) ◽  
pp. 649-656 ◽  
Author(s):  
Xiang-Yan Zhang ◽  
Yang Yu ◽  
Ping He ◽  
Yi-Xuan Zhang ◽  
Bao-Yu Hu ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Outer Membrane ◽  
Outer Membrane Proteins ◽  
Immunoblot Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Leptospira Interrogans ◽  
Genes Encoding ◽  
Leptospira Borgpetersenii ◽  
High Degree

AbstractLeptospiral outer membrane proteins (OMPs) are highly conserved in different species, and play an essential role in the development of new immunoprotection and serodiagnosis strategies. The genes encoding LipL21, LipL32 and OmpL1 were cloned from the complete genome sequence of Leptospira interrogans serovar lai strain Lai and expressed in vitro. Sequence comparison analysis revealed that the three genes were highly conserved among distinct epidemic leptospires, including three major epidemic species Leptospira interrogans, Leptospira borgpetersenii and Leptospira weilii, in China. Immunoblot analysis was further performed to scrutinize 15 epidemic Leptospira reference strains using the antisera of the recombinant OMPs. Both immunoblot assay and reverse transcription-polymerase chain reaction demonstrated that these three OMPs were conservatively expressed in pathogenic L. interrogans strains and other pathogenic leptospires. Additionally, the use of these recombinant OMPs as antigens in enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of leptospirosis was evaluated. The recombinant LipL32 and OmpL1 proteins showed a high degree of ELISA reactivity with sera from patients infected with L. interrogans strain Lai and other pathogenic leptospires. These results may contribute to the identification of candidates for broad-range vaccines and immunodiagnostic antigens in further research.

Sign in / Sign up

Export Citation Format

Share Document