scholarly journals Proteome Analysis of the Plasma Membrane ofMycobacterium tuberculosis

2002 ◽  
Vol 3 (6) ◽  
pp. 470-483 ◽  
Author(s):  
Sudhir Sinha ◽  
Shalini Arora ◽  
K. Kosalai ◽  
Abdelkader Namane ◽  
Alex S. Pym ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Drug Targets ◽  
Proteome Analysis ◽  
Membrane Vesicles ◽  
Genome Database ◽  
Peptide Mass ◽  
Proteome Database ◽  
Membrane Bound ◽  
Shock Proteins ◽  
Novel Drug

The plasmamembrane of Mycobacterium tuberculosisis likely to contain proteins that could serve as novel drug targets, diagnostic probes or even components of a vaccine against tuberculosis. With this in mind, we have undertaken proteome analysis of the membrane ofM. tuberculosisH37Rv. Isolated membrane vesicles were extracted with either a detergent (Triton X114) or an alkaline buffer (carbonate) following two of the protocols recommended for membrane protein enrichment. Proteins were resolved by 2D-GE using immobilized pH gradient (IPG) strips, and identified by peptide mass mapping utilizing theM. tuberculosisgenome database. The two extraction procedures yielded patterns with minimal overlap. Only two proteins, both HSPs, showed a common presence. MALDI–MS analysis of 61 spots led to the identification of 32 proteins, 17 of which were new to theM. tuberculosisproteome database. We classified 19 of the identified proteins as ‘membrane-associated’; 14 of these were further classified as ‘membrane-bound’, three of which were lipoproteins. The remaining proteins included four heat-shock proteins and several enzymes involved in energy or lipid metabolism. Extraction with Triton X114 was found to be more effective than carbonate for detecting ‘putative’M. tuberculosismembrane proteins. The protocol was also found to be suitable for comparing BCG andM. tuberculosismembranes, identifying ESAT-6 as being expressed selectively inM. tuberculosis. While this study demonstrates for the first time some of the membrane proteins ofM. tuberculosis, it also underscores the problems associated with proteomic analysis of a complex membrane such as that of a mycobacterium.

Renal neutral α-d-glucosidase has no role in transport of d-glucose derived from maltose hydrolysis

1998 ◽  
Vol 274 (4) ◽  
pp. R1150-R1157
Author(s):  
Jean Giudicelli ◽  
Pascale Delque-Bayer ◽  
Pierre Sudaka ◽  
Jean-Claude Poiree
Keyword(s):  
Membrane Proteins ◽  
Membrane Vesicles ◽  
Hydrolytic Activity ◽  
The Other ◽  
Transmembrane Transport ◽  
Artificial Membrane ◽  
Enzyme Localization ◽  
Free Glucose ◽  
Membrane Bound ◽  
Rule Out

To reinvestigate the “hydrolase-related transport” concept, neutral α-d-glucosidase, a membrane-bound disaccharidase of renal proximal tubule, was first purified from brush-border membranes and then asymmetrically reincorporated into egg phosphatidylcholine vesicles. Proteolytic treatments and immunotitration studies demonstrated that this enzyme was integrated in native and artificial membrane vesicles with a similar topology. The uptake of free glucose and glucose produced by maltose hydrolysis was studied using 1) proteoliposomes containing integrated neutral α-d-glucosidase, in combination with other membrane proteins, and 2) proteoliposomes containing only the other membrane proteins but incubated in a medium containing neutral α-d-glucosidase in its hydrophilic form. No modification was observed in the uptake of freed-glucose or d-glucose produced by maltose hydrolysis, regardless of enzyme localization. In contrast to previous findings on the hydrolase-related transport concept, these results rule out any participation of neutral α-d-glucosidase in the transport of free glucose or glucose produced by maltose hydrolysis. Hydrolytic activity and transmembrane transport appear to be two independent and sequential steps.

scholarly journals Proteins in the periplasmic space and outer membrane vesicles ofRhizobium etliCE3 grown in minimal medium are largely distinct and change with growth phase

2018 ◽  
Author(s):  
Hermenegildo Taboada ◽  
Niurka Meneses ◽  
Michael F. Dunn ◽  
Carmen Vargas-Lagunas ◽  
Natasha Buchs ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Minimal Medium ◽  
Proteome Analysis ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Periplasmic Space ◽  
Rhizobium Etli ◽  
Multidrug Efflux ◽  
Secretion Systems ◽  
Shock Proteins

ABSTRACTRhizobium etliCE3 grown in succinate-ammonium minimal medium (MM) excreted outer membrane vesicles (OMVs) with diameters of 40 to 100 nm. Proteins from the OMVs and the periplasmic space were isolated from 6 and 24 h cultures and identified by proteome analysis. A total 770 proteins were identified: 73.8 and 21.3 % of these proteins occurred only in the periplasm and OMVs, respectively, and only 4.9 % were found in both locations. The majority of proteins found in either location were present only at 6 or 24 h: in the periplasm and OMVs, only 24 and 9 % of proteins, respectively, were present at both sampling times, indicating a time-dependent differential sorting of proteins into the two compartments. The OMVs contained proteins with physiologically varied roles, includingRhizobiumadhering proteins (Rap), polysaccharidases, polysaccharide export proteins, autoaggregation and adherence proteins, glycosyl transferases, peptidoglycan binding and cross-linking enzymes, potential cell wall modifying enzymes, porins, multidrug efflux RND family proteins, ABC transporter proteins, and heat shock proteins. As expected, proteins with known periplasmic localizations (phosphatases, phosphodiesterases, pyrophosphatases) were found only in the periplasm, along with numerous proteins involved in amino acid and carbohydrate metabolism and transport. Nearly one-quarter of the proteins present in the OMVs were also found in our previous analysis of theR. etlitotal exproteome of MM-grown cells, indicating that these nanoparticles are an important mechanism for protein excretion in this species.IMPORTANCEThe reduction of atmospheric nitrogen to ammonia by rhizobia symbiotically associated with legumes is of major importance in sustainable agricultural. Rhizobia excrete a variety of symbiotically important proteins using canonical secretion systems. In this work, we show thatRhizobium etligrown in culture also excretes proteins in membrane-enclosed structures called outer membrane vesicles (OMVs). This study reports OMV production by rhizobia. Proteins identified in the OMVs included Rhizobium adhering (Rap) and autoaggregation proteins, polysaccharidases, RTX toxins, porins and multidrug efflux proteins. Some of these proteins have important roles in theR. etli-common bean symbiosis, and their packaging into OMVs could deliver them to the environment in a concentrated yet diffusible form protected from degradation. The work described here provides a basis for future studies on the function of rhizobial OMVs in free life and symbiosis.

scholarly journals Prediction of Novel Drug Targets and Vaccine Candidates against Human Lice (Insecta), Acari (Arachnida), and Their Associated Pathogens

Vaccines ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 8
Author(s):  
Abid Ali ◽  
Shabir Ahmad ◽  
Pedro Machado Medeiros de Albuquerque ◽  
Atif Kamil ◽  
Fahdah Ayed Alshammari ◽  
...  
Keyword(s):  
Metabolic Pathways ◽  
Drug Targets ◽  
Babesia Microti ◽  
Economic Losses ◽  
Borrelia Miyamotoi ◽  
Vaccine Candidates ◽  
Essential Proteins ◽  
Membrane Bound ◽  
Novel Drug ◽  
Novel Drug Targets

The emergence of drug-resistant lice, acari, and their associated pathogens (APs) is associated with economic losses; thus, it is essential to find new appropriate therapeutic approaches. In the present study, a subtractive proteomics approach was used to predict suitable therapeutics against these vectors and their infectious agents. We found 9701 proteins in the lice (Pediculus humanus var. corporis) and acari (Ixodes scapularis, Leptotrombidium deliense), and 4822 proteins in the proteomes of their APs (Babesia microti, Borreliella mayonii, Borrelia miyamotoi, Borrelia recurrentis, Rickettsia prowazekii, Orientia tsutsugamushi str. Boryong) that were non-homologous to host proteins. Among these non-homologous proteins, 365 proteins of lice and acari, and 630 proteins of APs, were predicted as essential proteins. Twelve unique essential proteins were predicted to be involved in four unique metabolic pathways of lice and acari, and 103 unique proteins were found to be involved in 75 unique metabolic pathways of APs. The sub cellular localization analysis of 115 unique essential proteins of lice and acari and their APs revealed that 61 proteins were cytoplasmic, 42 as membrane-bound proteins and 12 proteins with multiple localization. The druggability analysis of the identified 73 cytoplasmic and multiple localization essential proteins revealed 22 druggable targets and 51 novel drug targets that participate in unique pathways of lice and acari and their APs. Further, the predicted 42 membrane bound proteins could be potential vaccine candidates. Screening of useful inhibitors against these novel targets may result in finding novel compounds efficient for the control of these parasites.

Comparative Proteome Analysis of Mycobacterium Tuberculosis Strains - H37Ra, H37Rv, CCDC5180, and CAS/NITR204: A Step Forward to Identify Novel Drug Targets

2020 ◽  
Vol 17 (11) ◽  
pp. 1422-1431
Author(s):  
Shradheya R.R. Gupta ◽  
Ekta Gupta ◽  
Avnam Ohri ◽  
Sandeep Kumar Shrivastava ◽  
Sumita Kachhwaha ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Drug Target ◽  
Drug Targets ◽  
Virulent Strain ◽  
Proteome Analysis ◽  
Target Protein ◽  
Central Asian ◽  
Virulent Strains ◽  
Novel Drug ◽  
Novel Drug Targets

Background: Mycobacterium tuberculosis is a causative agent of tuberculosis. It is a non-motile, acid-fast, obligatory aerobic bacterium. Finding novel drug targets in Mycobacterium tuberculosis has become extremely important as the bacterium is evolving into a more dangerous multi-drug resistant pathogen. The predominant strains in India belong to the Central-Asian, East- African Indian, and Beijing clad. For the same reason, the whole proteomes of a non-virulent strain (H37Ra), a virulent (H37Rv) and two clinical strains, a Central-Asian clad (CAS/NITR204) and a Beijing clad (CCDC5180) have been selected for comparative study. Selecting a phylogenetically close and majorly studied non-virulent strain is helpful in removing the common and undesired proteins from the study. Objective: The study compares the whole proteome of non-virulent strain with the other three virulent strains to find a unique protein responsible for virulence in virulent strains. It is expected that the drugs developed against identified targets will be specific to the virulent strains. Additionally, to assure minimal toxicity to the host, we also screened the human proteome. Methods: Comparative proteome analysis was used for target identification and in silico validation of identified target protein Rv2466c, identification of the respective ligand of the identified target protein and binding interaction study using Molecular docking and Molecular Dynamic Simulation study were used in this study. Results and Discussion: Finally, eleven proteins were found to be unique in virulent strain only and out of which, Rv2466c (PDB-ID: 4ZIL) was found to be an essential protein and identified as a putative drug target protein for further study. The compound glutathione was found to be a suitable inhibitor for Rv2466c. In this study, we used a comparative proteomics approach to identify novel target proteins. Conclusion: This study is unique as we are assured that the study will move forward the research in a new direction to cure the deadly disease (tuberculosis) caused by Mycobacterium tuberculosis. Rv2466c was identified as a novel drug target and glutathione as a respective ligand of Rv2466c. Discovery of the novel drug target as well as the drug will provide a solution to drug resistance as well as the infection caused by Mycobacterium tuberculosis.

Novel drug targets in 2019

2020 ◽  
Vol 19 (5) ◽  
pp. 300-300 ◽  
Author(s):  
Sorin Avram ◽  
Liliana Halip ◽  
Ramona Curpan ◽  
Tudor I. Oprea
Keyword(s):  
Drug Targets ◽  
Novel Drug ◽  
Novel Drug Targets

Malaria Heat Shock Proteins: Drug Targets that Chaperone other Drug Targets

2010 ◽  
Vol 10 (3) ◽  
pp. 147-157 ◽  
Author(s):  
E.-R. Pesce ◽  
I.L. Cockburn ◽  
J.L. Goble ◽  
L.L. Stephens ◽  
G.L. Blatch
Keyword(s):  
Heat Shock ◽  
Heat Shock Proteins ◽  
Drug Targets ◽  
Shock Proteins
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