Recombinant Haemophilus ducreyi Lipid A export ATP-binding/permease protein MsbA (msbA)

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Cat. No.
BT2147206
Source
in vitro E.coli expression system
Synonyms
msbA; HD_1630; ATP-dependent lipid A-core flippase; Lipid A export ATP-binding/permease protein MsbA
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Description

Overview of MsbA in Haemophilus ducreyi

MsbA is an ATP-binding cassette (ABC) transporter responsible for flipping lipid A—a key component of lipopolysaccharides (LPS)—from the cytoplasmic to the periplasmic leaflet of the inner membrane . In Haemophilus ducreyi, this protein is encoded by the gene msbA (UniProt ID: Q7VL52) and is indispensable for bacterial viability .

Recombinant MsbA Production

Recombinant MsbA from H. ducreyi is expressed in Escherichia coli with an N-terminal His-tag for purification . Key specifications include:

ParameterDetails
Expression SystemE. coli
TagN-terminal His tag
Protein LengthFull-length (1–582 amino acids)
Purity>90% (verified by SDS-PAGE)
StorageLyophilized powder in Tris/PBS buffer with 6% trehalose (pH 8.0) at -80°C
ReconstitutionDeionized water with 50% glycerol recommended for stability

Mechanism of Action

MsbA operates via an ATP-dependent "trap and flip" mechanism:

  • Inward-facing conformation: Binds lipid A in the cytoplasmic leaflet .

  • ATP hydrolysis: Drives conformational changes to transport lipid A across the membrane .

  • Periplasmic release: Lipid A is transferred to the Lpt transport system for outer membrane assembly .

Antibiotic Development

MsbA’s essentiality in Gram-negative pathogens makes it a prime target for inhibitors. For example:

  • G907: A bactericidal antagonist that locks MsbA in an inward-facing conformation .

  • Lipid A analogs: Used to study substrate binding and transport kinetics .

Biochemical Assays

  • ATPase activity assays: Measure hydrolysis rates stimulated by lipid A or daunomycin .

  • Cross-linking studies: Identify conformational changes using tools like DARPin_55 .

Comparative Analysis with Homologs

SpeciesUniProt IDKey Differences
Pseudomonas fluorescensQ4KJB2Longer sequence (601 aa vs. 582 aa) and altered transmembrane helices
Escherichia coliP60752Higher ATPase activity and broader substrate specificity

Challenges and Future Directions

  • Structural dynamics: High-resolution cryo-EM or X-ray structures of H. ducreyi MsbA in multiple conformations are needed .

  • Drug specificity: Designing inhibitors that avoid off-target effects on human ABC transporters .

Product Specs

Form
Lyophilized powder
Note: While we prioritize shipping the format currently in stock, please specify your format preference in order notes for customized fulfillment.
Lead Time
Delivery times vary depending on the purchasing method and location. Please contact your local distributor for precise delivery estimates.
Note: Our standard shipping includes blue ice packs. Dry ice shipping requires prior arrangement and incurs additional charges.
Notes
Avoid repeated freeze-thaw cycles. Store working aliquots at 4°C for up to one week.
Reconstitution
Centrifuge the vial briefly before opening to consolidate the contents. Reconstitute the protein in sterile, deionized water to a concentration of 0.1-1.0 mg/mL. For long-term storage, we recommend adding 5-50% glycerol (final concentration) and aliquoting at -20°C/-80°C. Our standard glycerol concentration is 50%, which may serve as a guideline for your use.
Shelf Life
Shelf life depends on various factors including storage conditions, buffer composition, temperature, and the protein's inherent stability. Generally, liquid formulations have a 6-month shelf life at -20°C/-80°C, while lyophilized forms have a 12-month shelf life at -20°C/-80°C.
Storage Condition
Upon receipt, store at -20°C/-80°C. Aliquoting is essential for multiple uses. Avoid repeated freeze-thaw cycles.
Tag Info
The tag type is determined during the manufacturing process.
The tag type is determined during production. If you require a specific tag, please inform us, and we will prioritize its development.
Synonyms
msbA; HD_1630; ATP-dependent lipid A-core flippase; Lipid A export ATP-binding/permease protein MsbA
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Expression Region
1-582
Protein Length
full length protein
Species
Haemophilus ducreyi (strain 35000HP / ATCC 700724)
Target Names
msbA
Target Protein Sequence
MQETDLSIIKTFKRLFPIIRNYKWGLIAASVALILNALVDSSLIYLLKPLLDDGFGKADN AFLKQMAILVMLFILLRGVSNYIASYCLSWVSGKVVMTLRRNIFQHLMYMPVSYFDKNPT GRLLSRVTYDTEMVASSSSYVLVTIVREGAYLISLFAVMVYTSWQLSIVLFLLAPIIAFL ISIVSKRFRILSRNIQNSMGELTVTTEQMLKGHKVVLSFGGQKVEKARFDRVSNDMRRKG MKIVSADGISDGLVQLIASLALSAVLYVATFPEVMSENLTAGSFTVVFSSMMAMLRPLKS LTSVNSQFQRGMAACQTLFEFLDLKTEKNNGTKQVERAQGCITFDNVIFSYEGKEEQALN QVSFTIPQGKTVALVGRSGSGKSTIASLLTRFYDVNEGQILLDGVNIEEYTLENLREQCS VVSQQVHLFNDTIANNIAYAAKDKYSRAQIIAAAQAAHAMEFIEKLEQGLDTVIGENGAS LSGGQRQRLAIARALLRNAPVLVLDEATSALDTESELAIQSALAALQKNKTVLVIAHRLS TIEKADEILVVDQGKIVERGSHEQLLAKGGAYKQLYSMQFSE
Uniprot No.
Q7VL52

Target Background

Function

Recombinant Haemophilus ducreyi Lipid A export ATP-binding/permease protein MsbA (msbA) is involved in lipopolysaccharide (LPS) biosynthesis. It facilitates the translocation of lipid A-core from the inner to the outer leaflet of the inner membrane. Transmembrane domains (TMDs) form a pore within the inner membrane, while the ATP-binding domain (NBD) provides the necessary energy for this process.

Database Links

KEGG: hdu:HD_1630

STRING: 233412.HD1630

Protein Families
ABC transporter superfamily, Lipid exporter (TC 3.A.1.106) family
Subcellular Location
Cell inner membrane; Multi-pass membrane protein.

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