Recombinant Agrobacterium tumefaciens UPF0060 membrane protein Atu1058 (Atu1058)
Product Specs
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Target Background
KEGG: atu:Atu1058
STRING: 176299.Atu1058
Q&A
What is the functional role of Atu1058 in Agrobacterium tumefaciens?
Atu1058, a UPF0060 membrane protein (UniProt ID Q8UGH9), is annotated as a hypothetical protein with limited functional characterization. Current evidence suggests it is a full-length membrane protein (1–107 amino acids) expressed in A. tumefaciens, but its specific role in bacterial physiology or plant transformation remains unclear. Structural analysis reveals hydrophobic regions consistent with membrane localization, potentially implicating it in transmembrane signaling or substrate transport. Further studies are needed to elucidate its interaction with host plants or bacterial virulence factors .
How is the recombinant Atu1058 protein produced and purified?
Recombinant Atu1058 is typically expressed in E. coli with an N-terminal His-tag for affinity purification. Key steps include:
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Expression: Induction of E. coli cultures (e.g., BL21(DE3)) with IPTG under optimized conditions.
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Purification: Nickel-affinity chromatography to isolate the His-tagged protein, followed by SDS-PAGE validation (>90% purity) .
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Storage: Lyophilized powder stored at −20°C/−80°C in Tris/PBS buffer with 6% trehalose and 50% glycerol to prevent degradation .
| Parameter | Value/Method | Source |
|---|---|---|
| Expression Host | E. coli | |
| Fusion Tag | N-terminal His-tag | |
| Purification Method | Nickel-affinity chromatography | |
| Purity | >90% (SDS-PAGE) |
What challenges arise when using Atu1058 for plant transformation studies?
Key challenges include:
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Variable Transformation Efficiency: Atu1058’s role in Agrobacterium-plant interactions is poorly understood, making it hard to predict its impact on T-DNA transfer. Studies show strain-specific differences (e.g., Super-Agrobacterium ver. 4 enhances efficiency in tomato but not Nicotiana) .
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Protein Stability: Repeated freeze-thaw cycles degrade Atu1058, necessitating aliquoting and short-term storage at 4°C .
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Functional Redundancy: Overlapping roles with other membrane proteins may obscure its specific contribution to transformation.
What experimental approaches are effective for studying Atu1058’s role in genetic transformation?
How can researchers optimize the expression of recombinant Atu1058 in E. coli?
What data analysis methods are recommended for interpreting results involving Atu1058?
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Quantitative Analysis: Use ANOVA or t-tests to compare transformation efficiencies between strains (e.g., wild-type vs. acdS/ gabT-engineered Agrobacterium) .
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Proteomic Data: Apply label-free quantification (LFQ) or SILAC for LC-MS/MS data to identify Atu1058-specific interactors .
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Structural Modeling: Predict transmembrane domains using tools like Phyre2 or TMHMM.
How does Atu1058 compare to other UPF0060 membrane proteins in function?
UPF0060 proteins are poorly characterized, but Atu1058’s His-tagged recombinant form enables functional studies. Unlike many hypothetical proteins, Atu1058 is expressed at detectable levels in E. coli, making it tractable for biochemical assays . Comparative genomics could identify conserved motifs or orthologs in related Agrobacterium species.
