Recombinant Mycoplasma pneumoniae Uncharacterized protein MPN_312 (MPN_312)
Description
Molecular Characterization of Recombinant MPN_312
MPN_312 (UniProt ID: P75450) is encoded by the MPN_312 gene and classified as a terminal organelle-associated protein. Recombinant forms are produced in Escherichia coli with a hexahistidine (His) tag for purification .
Biological Role in M. pneumoniae Pathogenesis
MPN_312 (identified as P24 in functional studies) is essential for terminal organelle assembly, gliding motility, and cell division .
Functional Insights:
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Terminal Organelle Assembly:
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Gliding Motility Regulation:
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Interaction Network:
Expression and Purification Protocol111:
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Host System: E. coli (BL21 or similar strains).
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Tags: N-terminal His tag (± C-terminal Myc tag for detection) .
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Storage: Lyophilized in Tris/PBS buffer with 6% trehalose (pH 8.0); stable at -80°C .
Research Applications:
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Mechanistic Studies:
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Antigenic Analysis:
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Vaccine Development:
Evolutionary and Clinical Relevance
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Genetic Stability:
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Antibiotic Resistance Context:
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Epidemiological Trends:
Challenges and Future Directions
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Structural Resolution: No crystal structure available; computational modeling needed to map interaction interfaces .
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Functional Redundancy: Overlap with P41 complicates disentangling individual roles in motility .
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Therapeutic Potential: Indirect targeting via terminal organelle disruption remains unexplored .
Product Specs
Q&A
Basic Research Questions
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What is the MPN_312 gene and what protein does it encode in Mycoplasma pneumoniae?
MPN_312 encodes the protein P24, a terminal organelle protein in Mycoplasma pneumoniae. P24 is a cell division-associated protein that plays a critical role in terminal organelle development and function. Research by Hasselbring and Krause (2007) established that P24 is required for normal initiation of terminal organelle formation . Structurally, P24 localizes to the proximal end of the terminal organelle in wild-type cells, similar to P41, which is predicted to be translationally coupled to P24 .
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What experimental methods are commonly used to study MPN_312/P24 function?
Several complementary approaches are employed to study P24:
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Transposon mutagenesis: Disruption of MPN312 via transposon insertion to generate loss-of-function mutants
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Fluorescent protein fusions: Creating P24-YFP (Yellow Fluorescent Protein) fusion proteins to track localization in living cells
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Western immunoblotting: Quantifying steady-state levels of P24 in wild-type and mutant strains
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Time-lapse imaging: Evaluating terminal organelle development and motility
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Complementation studies: Reintroducing wild-type MPN312 allele via transposon delivery to confirm phenotype rescue
These methods allow researchers to characterize the spatiotemporal dynamics and functional significance of P24 in M. pneumoniae biology.
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How does P24 contribute to the structure and function of M. pneumoniae?
P24 plays a crucial role in M. pneumoniae cellular function through multiple mechanisms:
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Terminal organelle development: P24 is required for normal initiation and positioning of the terminal organelle development
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Cell division: Loss of P24 results in delayed displacement of the terminal organelle to the opposite pole during cell division
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Cellular motility: P24 mutants show delayed reacquisition of gliding motility following cell division
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Functional coupling: P24 is functionally associated with P41, as they appear to be translationally coupled, though P41 remains stable even when P24 levels are reduced
The terminal organelle is essential for M. pneumoniae pathogenicity, as it mediates both cytadherence to host respiratory epithelium and gliding motility, making P24 indirectly important for virulence.
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