Recombinant Chlamydomonas reinhardtii ATP synthase subunit b, chloroplastic (atpF)
Description
Introduction to Recombinant Chlamydomonas reinhardtii ATP Synthase Subunit b (atpF)
The recombinant Chlamydomonas reinhardtii ATP synthase subunit b (atpF) is a chloroplast-encoded protein critical for the biogenesis and function of the ATP synthase complex in photosynthetic organisms. This subunit, part of the peripheral stalk of the Fo sector, facilitates proton translocation and stabilizes the ATP synthase structure. The recombinant version (Cat.No. RFL35034CF) is produced in Escherichia coli with an N-terminal His-tag for purification and functional studies .
Role in ATP Synthase Biogenesis and Function
The atpF subunit (b) pairs with ATPG-encoded subunit b' in the peripheral stalk, forming a critical structural and functional bridge between the Fo and F1 sectors of the ATP synthase complex .
Functional Significance
-
Proton Translocation: Facilitates proton flow through the Fo sector, driving ATP synthesis in chloroplasts .
-
Stability: Mutants in atpF or ATPG genes (e.g., frame-shift or CRISPR knockout) result in complete loss of ATP synthase function and accumulation .
-
Interactions: Coordinated biogenesis with subunits like atpE (ε-chain) requires nuclear-encoded factors (e.g., MDE1, an octotricopeptide repeat protein) to stabilize mRNA and enable translation .
Biosynthesis and Purification
| Step | Detail | Source |
|---|---|---|
| Host Organism | E. coli (recombinant expression) | |
| Purification | Nickel affinity chromatography (His-tag) | |
| Yield | Not explicitly reported; typical E. coli yields vary by construct |
Research Applications
-
Structural Studies: His-tag enables immobilization for crystallography or cryo-EM .
-
Enzyme Activity Assays: Functional testing of ATP synthase subunit interactions .
-
Biotechnological Tools: Potential use in biofuel production or artificial photosynthesis systems .
Limitations
-
Expression Heterogeneity: While E. coli avoids the low transgene expression issues seen in Chlamydomonas , proper folding and solubility remain critical .
-
Storage Sensitivity: Repeated freeze-thaw cycles degrade activity; aliquoting for long-term storage at -20°C/-80°C is essential .
Key Research Findings
-
Peripheral Stalk Dynamics: Mutants in atpF or ATPG disrupt ATP synthase assembly, highlighting the subunits’ role in stabilizing the Fo-F1 interface .
-
Posttranscriptional Regulation: MDE1 stabilizes atpE mRNA, ensuring coordinated synthesis of ATP synthase subunits .
Comparative Analysis of ATP Synthase Subunits
Product Specs
Note: While we prioritize shipping the format currently in stock, please specify your preferred format in order notes for customized fulfillment.
Note: Standard shipping includes blue ice packs. Dry ice shipping requires prior arrangement and incurs additional charges.
The specific tag will be determined during production. If you require a specific tag, please inform us; we will prioritize its inclusion.
Target Background
F1F0 ATP synthase synthesizes ATP from ADP using a proton or sodium gradient. This enzyme comprises two domains: F1, the extramembranous catalytic core, and F0, the membrane proton channel. These domains are linked by a central and peripheral stalk. ATP synthesis in the F1 catalytic domain is coupled to proton translocation through a rotary mechanism involving the central stalk subunits. This protein is a component of the F0 channel and forms part of the peripheral stalk, connecting F1 to F0.
KEGG: cre:ChreCp054
STRING: 3055.DAA00955
