Recombinant Mycoplasma genitalium Putative ABC transporter ATP-binding protein MG015 (MG015)
Description
Functional Role in M. genitalium
MG015 is implicated in:
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Multidrug Resistance (MDR): ABC transporters like MG015 efflux antimicrobial agents (e.g., fluoroquinolones, ethidium bromide) via ATP-driven conformational changes .
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Pathogen Survival: Overexpression of homologous genes (md1/md2 in M. hominis) correlates with reduced drug uptake and increased MICs .
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Operon Organization: mg015 overlaps with mg014 by 8 nucleotides, suggesting coordinated regulation .
Recombinant Production and Applications
Recombinant MG015 is commercially produced for research purposes. Key details include:
| Parameter | Specification |
|---|---|
| Host System | E. coli |
| Tag | N-terminal His tag |
| Applications | - Mechanistic studies of ABC transporters - Antibiotic resistance screening |
4.1. Expression and Overexpression
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In M. hominis, homologous genes (md1/md2) showed 2–10× increased expression in ethidium bromide-resistant strains, directly linked to reduced ciprofloxacin uptake .
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MG015’s ATPase activity drives substrate translocation via a "power stroke" mechanism, involving ATP binding, NBD dimerization, and transmembrane helix reorientation .
Implications for Antimicrobial Resistance
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Clinical Relevance: Overexpression of ABC transporters like MG015 complicates treatment of M. genitalium infections, particularly with rising macrolide and fluoroquinolone resistance .
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Diagnostic Targets: Genetic variation in ABC transporter operons (e.g., MgPa operon) necessitates NAAT-based detection to avoid false-negative results in PCR assays .
Future Research Directions
Product Specs
Note: While we prioritize shipping the format currently in stock, we are happy to accommodate specific format requests. Please indicate your preferred format in the order notes, and we will fulfill your requirement.
Note: All proteins are shipped with standard blue ice packs. For shipments requiring dry ice, please contact us in advance. Additional fees may apply.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. For lyophilized form, the shelf life is 12 months at -20°C/-80°C.
Target Background
KEGG: mge:MG_015
STRING: 243273.MgenG_010200002816
Q&A
Basic Research Questions
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What is MG015 and what is its functional role in Mycoplasma genitalium?
MG015 is a putative ABC transporter ATP-binding protein found in Mycoplasma genitalium (strain ATCC 33530 / G-37 / NCTC 10195). It belongs to the ABC transporter superfamily . ABC transporters are primary active transporters found in all kingdoms of life and are involved in the transport of various substrates across membranes .
While the specific substrates transported by MG015 are not definitively characterized, genomic analyses have identified it as a putative multidrug resistance (MDR) gene, similar to MG014 . Studies in related Mycoplasma species suggest that these proteins may be involved in antimicrobial resistance mechanisms.
The protein consists of 589 amino acids and has a molecular mass of approximately 66.1 kDa . As an ABC transporter ATP-binding protein, MG015 likely couples ATP binding and hydrolysis with conformational changes that facilitate substrate transport across the membrane.
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How is recombinant MG015 typically produced for research applications?
Recombinant MG015 is typically produced using heterologous expression systems, with E. coli being the most common host organism . The production process generally involves:
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Cloning the MG015 gene into an expression vector (often with an N-terminal His-tag for purification)
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Transformation into an E. coli expression strain (commonly BL21-DE3)
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Induction of protein expression
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Cell lysis and protein extraction
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Purification via affinity chromatography using the His-tag
Similar to other membrane-associated proteins, MG015 can form inclusion bodies requiring denaturing conditions for solubilization . Purification protocols often include:
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Initial purification under denaturing conditions with 6-8M urea
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Gradual refolding through dialysis with decreasing urea concentrations
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Final purification steps to remove misfolded protein and contaminants
Storage recommendations typically include adding glycerol (often 50%) and storing at -20°C or -80°C to maintain stability .
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What experimental approaches are used to study MG015 interaction with other proteins?
Several experimental approaches can be employed to study MG015 interactions:
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Pull-down assays: Using His-tagged recombinant MG015 to identify binding partners from cell lysates
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Yeast two-hybrid screening: For detecting protein-protein interactions in vivo
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Surface plasmon resonance (SPR): For measuring binding kinetics and affinity
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Co-immunoprecipitation: To verify interactions in relevant cellular contexts
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Crosslinking studies: To capture transient interactions
These techniques have been applied to study ABC transporters in general and can be adapted specifically for MG015. For example, similar approaches have been used to study interactions between Mycoplasma genitalium proteins and host factors, as demonstrated in the study of MgPa interactions with host RPL35 .
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