Recombinant Escherichia coli Inner membrane transport permease ybhR (ybhR)

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Cat. No.

BT2125436

Source

in vitro E.coli expression system

Synonyms

ybhR; b0792; JW5803; Probable multidrug ABC transporter permease YbhR

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Description

Biochemical Properties

Gene Name: ybhR (synonyms: b0792, JW5803) .
UniProt ID: P0AFP9 (E. coli strain) , P0AFQ1 (Shigella flexneri homolog) .
Protein Length: 368 amino acids .
Domains:
- Transmembrane domain (TMD) with 6–12 membrane-spanning helices .
- Functional collaboration with YbhF (ATP-binding domain) and YbhS (membrane subunit) in the YbhFSR complex .

YbhR operates within the YbhFSR transporter complex, which demonstrates dual functionality:

Drug Efflux Activity

Substrates: Tetracycline, oxytetracycline, chlortetracycline, doxycycline, ethidium bromide (EB), and Hoechst33342 .
Mechanism: ATP hydrolysis by YbhF (NatA motif-containing ATPase) energizes substrate translocation across the membrane .
Experimental Evidence:
- Knockout strains of ybhF or ybhR show reduced efflux capacity, confirmed via ethidium bromide accumulation assays .
- Minimum inhibitory concentration (MIC) assays revealed hypersensitivity of knockout strains to tetracyclines .

Ion Transport Activity

Role: YbhFSR acts as a Na⁺(Li⁺)/H⁺ antiporter, enabling bacterial tolerance to high salinity and alkaline pH .
Key Motif: The NatA domain (residues 334–574 in YbhF) is critical for sodium transport .
Functional Validation:
- Heterologous expression of ybhF in Na⁺/H⁺ transporter-deficient E. coli KNabc restored growth under high NaCl/LiCl conditions .

Key Studies

Dual-Function Characterization (2020)
- Demonstrated YbhFSR’s role in both antibiotic resistance and ion homeostasis using MIC assays, fluorescence-based efflux measurements, and ATPase activity tests .
- Identified tetracyclines as primary substrates through competitive efflux inhibition .
Structural Homology Analysis
- Phylogenetic analysis revealed 31.3% sequence identity between YbhF and RB6469, a sodium transporter from Rhodopirellula baltica .
- Homology modeling predicted conserved ATP-binding motifs (Walker A/B, ABC signature) in YbhF .

Biotechnological Relevance

Drug Resistance Studies: YbhR is a target for understanding multidrug resistance mechanisms in Enterobacteriaceae .
Protein Engineering: Recombinant YbhR serves as a template for structure-function studies of ABC transporters .

Substrate Specificity of YbhFSR

Substrate	Efflux Activity	MIC Reduction in ΔybhFSR
Tetracycline	Yes	8-fold
Ethidium Bromide	Yes	4-fold
NaCl (100 mM)	N/A	Growth restoration

Comparison of Recombinant YbhR Variants

Feature	E. coli YbhR	Shigella YbhR
Host Species	E. coli K-12	Shigella flexneri 2a
UniProt Entry	P0AFP9	P0AFQ1
Applications	Efflux assays, structural studies	Comparative genomics

Product Specs

Form

Lyophilized powder
Note: We prioritize shipping the format currently in stock. However, if you have specific format requirements, please specify them when placing your order, and we will accommodate your request.

Lead Time

Delivery time may vary depending on the purchasing method and location. Please consult your local distributors for specific delivery timelines.
Note: All our proteins are shipped with standard blue ice packs. If you require dry ice shipping, please inform us in advance, as additional fees will apply.

Notes

Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Reconstitution

We recommend briefly centrifuging the vial before opening to ensure the contents settle at the bottom. Reconstitute the protein in deionized sterile 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 the solution at -20°C/-80°C. Our standard final glycerol concentration is 50%, which can serve as a reference.

Shelf Life

The shelf life of our products depends on several factors, including storage conditions, buffer components, storage temperature, and the inherent stability of the protein itself.
Generally, the shelf life for the liquid form is 6 months at -20°C/-80°C. Lyophilized form has a shelf life of 12 months at -20°C/-80°C.

Storage Condition

Store at -20°C/-80°C upon receipt. Aliquoting is necessary for multiple uses. Avoid repeated freeze-thaw cycles.

Tag Info

Tag type is determined during the manufacturing process.

Tag type is established during production. If you have a specific tag type requirement, please inform us, and we will prioritize developing the specified tag.

Synonyms

ybhR; b0792; JW5803; Probable multidrug ABC transporter permease YbhR

Buffer Before Lyophilization

Tris/PBS-based buffer, 6% Trehalose.

Datasheet

Please contact us to get it.

Expression Region

1-368

Protein Length

full length protein

Species

Escherichia coli (strain K12)

Target Names

ybhR

Target Protein Sequence

MFHRLWTLIRKELQSLLREPQTRAILILPVLIQVILFPFAATLEVTNATIAIYDEDNGEH SVELTQRFARASAFTHVLLLKSPQEIRPTIDTQKALLLVRFPADFSRKLDTFQTAPLQLI LDGRNSNSAQIAANYLQQIVKNYQQELLEGKPKPNNSELVVRNWYNPNLDYKWFVVPSLI AMITTIGVMIVTSLSVAREREQGTLDQLLVSPLTTWQIFIGKAVPALIVATFQATIVLAI GIWAYQIPFAGSLALFYFTMVIYGLSLVGFGLLISSLCSTQQQAFIGVFVFMMPAILLSG YVSPVENMPVWLQNLTWINPIRHFTDITKQIYLKDASLDIVWNSLWPLLVITATTGSAAY AMFRRKVM

Uniprot No.

P0AFP9

Target Background

Function

Recombinant Escherichia coli Inner membrane transport permease ybhR (ybhR) is a component of the ABC transporter complex YbhFSR. It is likely involved in the efflux of cefoperazone and may play a role in substrate translocation across the membrane.

Database Links

KEGG: ecj:JW5803

STRING: 316385.ECDH10B_0860

Protein Families

ABC-2 integral membrane protein family

Subcellular Location

Cell inner membrane; Multi-pass membrane protein.

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Recombinant Escherichia coli Inner membrane transport permease ybhR (ybhR)

Description

Biochemical Properties

Drug Efflux Activity

Ion Transport Activity

Key Studies

Biotechnological Relevance

Substrate Specificity of YbhFSR

Comparison of Recombinant YbhR Variants

Product Specs

Target Background

Quick Inquiry

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