Cloning of genes and characterization of immunogenic proteins of the antigenic mannoprotein superfamily in Aspergillus

AbstractMycobacterium abscessus (MAB) is a widely disseminated pathogenic non-tuberculous mycobacterium (NTM). Like with M. tuberculosis complex (MTBC), excreted / secreted (ES) proteins play an essential role for its virulence and survival inside the host. ES proteins contain highly immunogenic proteins, which are of interest for novel diagnostic assays and vaccines. Here, we used a robust bioinformatics pipeline to predict the secretome of the M. abscessus ATCC 19977 reference strain and fifteen clinical isolates belonging to all three MAB subspecies, M. abscessus subsp. abscessus, M. abscessus subsp. bolletii, and M. abscessus subsp. massiliense. We found that ~18% of the proteins encoded in the MAB genomes were predicted as secreted and that the three MAB subspecies shared > 85 % of the predicted secretomes. MAB isolates with a rough (R) colony morphotype showed larger predicted secretomes than isolates with a smooth (S) morphotype. Additionally, proteins exclusive to the secretomes of MAB R variants had higher antigenic densities than those exclusive to S variants, independently of the subspecies. For all investigated isolates, ES proteins had a significantly higher antigenic density than non-ES proteins. We identified 337 MAB ES proteins with homologues in previously investigated M. tuberculosis secretomes. Among these, 222 have previous experimental support of secretion, and some proteins showed homology with protein drug targets reported in the DrugBank database. The predicted MAB secretomes showed a higher abundance of proteins related to quorum-sensing and Mce domains as compared to MTBC indicating the importance of these pathways for MAB pathogenicity and virulence. Comparison of the predicted secretome of M. abscessus ATCC 19977 with the list of essential genes revealed that 99 secreted proteins corresponded to essential proteins required for in vitro growth. All predicted secretomes were deposited in the Secret-AAR web-server (http://microbiomics.ibt.unam.mx/tools/aar/index.php).

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Identification and Immunological Characterization of Somatic Proteins from Adults of Toxocara cati by Proteomics Technique

Iranian Journal of Parasitology ◽

10.18502/ijpa.v16i1.5508 ◽

2021 ◽

Author(s):

Nooshinmehr SOLEYMANI ◽

Ruth Birner GRUNBERGER ◽

Khalil ABNOUS ◽

Hassan BORJI ◽

Faezeh VAHDATI

Keyword(s):

Drug Targets ◽

Immunoblot Analysis ◽

Toxocara Cati ◽

Additional Information ◽

Immunogenic Proteins ◽

Etiological Agents ◽

Paratenic Hosts ◽

Medical University ◽

Somatic Antigens

Background: Toxocara cati is considered as one of the main etiological agents of toxocariasis with global and regional importance. As there is no information on proteomics of T. cati, herein, we reported the results obtained by proteomic analysis of somatic proteins extract, using a mass spectrometry (LC–MS/MS) approach. Methods: Somatic extract fractions were separated by two-dimensional SDSPAGE and were electro blotted on to PVDF membranes for immunoblot analysis, then collected the immunogenic spots which response of antibodies of the paratenic hosts (mice) to the antigens ( Mashhad, 2017), and analyzed by LC–MS/MS. The LC-MS/MS data were analyzed by Mascot database, Taxonomy Toxocara, and common contaminants, in Omics Center, Biotechnology Medical University of Graz (Austria, 2018). Result: The protein spots were isolated between 15–140 kDa ranges using 3–10 non-linear IPG strips and Brilliant Blue Coomassie. Ten proteins were characterized as immunogenic proteins, seven of them were identified and three of them were unknown proteins. Conclusion: This study provided additional information about the somatic antigens of T. cati, which can lead to the development of new strategies for novel immuno-modulators, drug targets, subunit vaccines and immunodiagnostic kits for toxocariasis.

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Identification and Characterization of Novel Immunogenic Proteins ofStreptococcus suisSerotype 2

Journal of Proteome Research ◽

10.1021/pr800196v ◽

2008 ◽

Vol 7 (9) ◽

pp. 4132-4142 ◽

Cited By ~ 48

Author(s):

Hongran Geng ◽

Li Zhu ◽

Yuan Yuan ◽

Wei Zhang ◽

Wenjun Li ◽

...

Keyword(s):

Immunogenic Proteins ◽

Identification And Characterization

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Characterization of immunogenic proteins of Trypanosoma evansi isolated from three different Indian hosts using hyperimmune sera and immune sera

Research in Veterinary Science ◽

10.1016/j.rvsc.2008.02.011 ◽

2008 ◽

Vol 85 (3) ◽

pp. 534-539 ◽

Cited By ~ 4

Author(s):

R. Laha ◽

N.K. Sasmal

Keyword(s):

Trypanosoma Evansi ◽

Immunogenic Proteins

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Characterization of lipid-modified immunogenic proteins of Treponema pallidum expressed in Escherichia coli

Microbial Pathogenesis ◽

10.1016/0882-4010(89)90053-3 ◽

1989 ◽

Vol 7 (3) ◽

pp. 175-188 ◽

Cited By ~ 14

Author(s):

Leo M Schouls ◽

Robert Mout ◽

Jan Dekker ◽

Jan D.A van Embden

Keyword(s):

Escherichia Coli ◽

Treponema Pallidum ◽

Immunogenic Proteins

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Characterization of Anti-Salmonella enterica Serotype Typhi Antibody Responses in Bacteremic Bangladeshi Patients by an Immunoaffinity Proteomics-Based Technology

Clinical and Vaccine Immunology ◽

10.1128/cvi.00104-10 ◽

2010 ◽

Vol 17 (8) ◽

pp. 1188-1195 ◽

Cited By ~ 30

Author(s):

Richelle C. Charles ◽

Alaullah Sheikh ◽

Bryan Krastins ◽

Jason B. Harris ◽

M. Saruar Bhuiyan ◽

...

Keyword(s):

Typhoid Fever ◽

Salmonella Enterica ◽

Iron Homeostasis ◽

Human Infection ◽

Diagnostic Assays ◽

Metabolic Functions ◽

Iga Antibody ◽

Immunogenic Proteins ◽

Typhoid Vaccines

ABSTRACT Salmonella enterica serotype Typhi is the cause of typhoid fever and a human-restricted pathogen. Currently available typhoid vaccines provide 50 to 90% protection for 2 to 5 years, and available practical diagnostic assays to identify individuals with typhoid fever lack sensitivity and/or specificity. Identifying immunogenic S. Typhi antigens expressed during human infection could lead to improved diagnostic assays and vaccines. Here we describe a platform immunoaffinity proteomics-based technology (IPT) that involves the use of columns charged with IgG, IgM, or IgA antibody fractions recovered from humans bacteremic with S. Typhi to capture S. Typhi proteins that were subsequently identified by mass spectrometry. This screening tool identifies immunogenic proteins recognized by antibodies from infected hosts. Using this technology and the plasma of patients with S. Typhi bacteremia in Bangladesh, we identified 57 proteins of S. Typhi, including proteins known to be immunogenic (PagC, HlyE, OmpA, and GroEL) and a number of proteins present in the human-restricted serotypes S. Typhi and S. Paratyphi A but rarely found in broader-host-range Salmonella spp. (HlyE, CdtB, PltA, and STY1364). We categorized identified proteins into a number of major groupings, including those involved in energy metabolism, protein synthesis, iron homeostasis, and biosynthetic and metabolic functions and those predicted to localize to the outer membrane. We assessed systemic and mucosal anti-HlyE responses in S. Typhi-infected patients and detected anti-HlyE responses at the time of clinical presentation in patients but not in controls. These findings could assist in the development of improved diagnostic assays.

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Morphological Characterization of Mycobacteriophage R1

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100051281 ◽

1974 ◽

Vol 32 ◽

pp. 254-255

Author(s):

B. L. Soloff ◽

T. A. Rado

Keyword(s):

Carbon Source ◽

Stationary Phase ◽

Growth Phase ◽

Minimal Medium ◽

Morphological Characterization ◽

Logarithmic Growth Phase ◽

Complete Lysis ◽

Lysogenic Culture ◽

Logarithmic Growth

Mycobacteriophage R1 was originally isolated from a lysogenic culture of M. butyricum. The virus was propagated on a leucine-requiring derivative of M. smegmatis, 607 leu−, isolated by nitrosoguanidine mutagenesis of typestrain ATCC 607. Growth was accomplished in a minimal medium containing glycerol and glucose as carbon source and enriched by the addition of 80 μg/ ml L-leucine. Bacteria in early logarithmic growth phase were infected with virus at a multiplicity of 5, and incubated with aeration for 8 hours. The partially lysed suspension was diluted 1:10 in growth medium and incubated for a further 8 hours. This permitted stationary phase cells to re-enter logarithmic growth and resulted in complete lysis of the culture.

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