Recombinant Mannheimia succiniciproducens Cell division protein ZapB (zapB)
Description
Role in Cell Division
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Z-Ring Formation ZapB is crucial for Z-ring formation, a key step in bacterial cell division .
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Divisome Recruitment ZapB is recruited to the divisome, the protein complex that orchestrates cell division, through direct interaction with ZapA .
ZapB Analogs in Other Bacteria
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ZapT in Caulobacter crescentus In Caulobacter crescentus, a protein called ZapT appears to play a similar role to ZapB in linking the divisome to the chromosome terminus . Unlike ZapB, ZapT is conserved in a broader range of bacteria beyond enteric bacteria .
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Interaction with ZapA ZapT interacts with ZapA and ZauP in C. crescentus, analogous to the ZapA-ZapB interaction in E. coli . This interaction is important for localizing ZapA to the cell pole and facilitating Z-ring formation .
Metabolic Engineering of Mannheimia succiniciproducens
Mannheimia succiniciproducens is a bacterium that can produce succinic acid . Metabolic engineering has been used to enhance succinic acid production by this organism .
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Succinic Acid Production M. succiniciproducens can produce a large amount of succinic acid under anaerobic conditions with CO$$_2$$ .
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CO$$_2$$ Fixation Phosphoenolpyruvate (PEP) carboxylation is a major CO$$_2$$-fixing step that directly impacts succinic acid production .
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Metabolic Pathways Key metabolic pathways leading to by-product formation have been removed to improve succinic acid production .
Effects of CO$$_2$$ on Mannheimia succiniciproducens Growth
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CO$$_2$$ Dependence M. succiniciproducens requires CO$$_2$$ for efficient growth and succinic acid production .
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Growth Suppression Cell growth is severely suppressed at low dissolved CO$$_2$$ concentrations .
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Enzyme Activity PEP carboxykinase (PckA) is crucial for succinic acid production and growth by providing additional ATP .
Overexpression of ZapT
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Cell Division Blockage Overexpression of ZapT in Caulobacter crescentus blocks cell division, leading to elongated cells and an increased fraction of cells with two chromosomes .
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MipZ Mislocalization Excess ZapT can interfere with the localization of the FtsZ inhibitor MipZ, potentially disrupting the normal cell division process .
Potential Role of ZapT in Spatial Coordination
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DNA-ZapT Complexes It is proposed that DNA-ZapT complexes may play a role in coordinating the terminus and divisome .
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Analogy to MatP-ZapB Similar to the MatP-ZapB interaction in E. coli, ZapT may form a complex with the terminus region of the chromosome and divisome components to recruit the Z-ring .
Product Specs
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Target Background
Function: A non-essential, abundant cell division factor crucial for proper Z-ring formation. It is recruited early to the divisome through direct interaction with FtsZ, stimulating Z-ring assembly and accelerating cell division within the cell cycle. Its recruitment to the Z-ring necessitates functional FtsA or ZipA.
KEGG: msu:MS2253
STRING: 221988.MS2253
Q&A
Basic Research Questions
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What is Mannheimia succiniciproducens and why is it significant in research?
Mannheimia succiniciproducens MBEL55E is a capnophilic gram-negative bacterium isolated from bovine rumen that efficiently produces succinic acid as a major fermentation product under anaerobic conditions in the presence of CO2 . Its significance in research stems from its ability to produce large amounts of succinic acid, a four-carbon dicarboxylic acid with industrial applications as a platform chemical for the manufacture of synthetic resins, biodegradable polymers, and chemical intermediates . The complete genome sequence of M. succiniciproducens has enabled comprehensive metabolic engineering to optimize succinic acid production .
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What is ZapB and what role does it play in bacterial cell division?
ZapB is a non-essential but abundant cell division factor required for proper Z-ring formation in bacteria . It is recruited early to the divisome through direct interaction with FtsZ, where it stimulates Z-ring assembly, thereby promoting cell division earlier in the cell cycle . The crystal structure reveals that ZapB exists as a dimer that is 100% coiled-coil, and in vitro, it self-assembles into long filaments and bundles . Its recruitment to the Z-ring requires functional FtsA or ZipA, and deletion of zapB results in delayed cell division and formation of ectopic Z rings and spirals, while overexpression leads to nucleoid condensation and aberrant cell divisions .
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What are the structural characteristics of M. succiniciproducens ZapB?
In Mannheimia succiniciproducens strain MBEL55E, ZapB is a 72 amino acid protein with a molecular mass of 8.4 kDa . Its amino acid sequence is MSLEILDQLEGKIKQAVETIQLLQLEVEELKEKNQQAQQANDELRSENEQLKGEHNNWQERLRSLLGQIDNV . The protein belongs to the ZapB family and exists as a coiled-coil dimer that can self-assemble into larger structures, which is critical for its function in Z-ring formation .
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How is recombinant M. succiniciproducens ZapB protein typically expressed and purified?
Recombinant M. succiniciproducens ZapB can be expressed in various expression systems, with mammalian cell systems being one documented approach . The expression typically covers the full-length protein or specific regions (such as 1-107) . For purification, the protein is often tagged (tag type determined during manufacturing) and can be purified using standard chromatography techniques to achieve >85% purity as verified by SDS-PAGE . After purification, the protein can be reconstituted in deionized sterile water to a concentration of 0.1-1.0 mg/mL, with glycerol (5-50% final concentration) added for long-term storage at -20°C/-80°C .
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What experimental methods are used to study ZapB localization and interactions?
Several methodological approaches are used to study ZapB localization and interactions:
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Bacterial two-hybrid analysis to detect protein-protein interactions, particularly with FtsZ
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Time-lapse microscopy using ZapB-GFP fusion proteins to visualize its localization at mid-cell in patterns similar to FtsZ
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Gene deletion studies to observe phenotypic changes such as delayed cell division and formation of ectopic Z rings
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Overexpression studies to observe effects like nucleoid condensation and aberrant cell divisions
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Immunofluorescence assays (IFA) for co-localization studies with other cell division proteins
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