Recombinant Escherichia coli Ferredoxin-type protein napH (napH)
Description
Definition and Biological Context
Recombinant Escherichia coli Ferredoxin-type protein NapH (NapH) is a genetically engineered iron-sulfur protein expressed in E. coli for research applications. NapH belongs to the ferredoxin family, which facilitates electron transfer in redox reactions. In E. coli, the napH gene (synonym: yejZ) encodes this protein, which is implicated in nitrate reduction pathways and redox homeostasis .
Expression and Purification
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Host System: Expressed in E. coli (strain K12) using vectors like p delta blue under bacteriophage lambda promoters .
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Solubility: Predominantly found in inclusion bodies, with partial solubility in the cytoplasmic fraction .
| Parameter | Details |
|---|---|
| Gene Name | napH (also yejZ) |
| UniProt ID | P33934 (E. coli strain K12) |
| Protein Length | 287 amino acids |
| Tag | N-terminal His tag |
| Expression Host | Escherichia coli |
Catalytic Activity
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Electron Transfer: NapH likely participates in electron shuttling, analogous to other ferredoxins, though its specific substrates in E. coli remain under investigation .
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Interaction Partners: Associates with redox enzymes such as nitrate reductase (Nap complex) .
Research Applications
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Enzyme Kinetics: Used to study electron transfer mechanisms in bacterial nitrate respiration .
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Structural Studies: Purified recombinant NapH enables X-ray crystallography and circular dichroism analyses .
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Oxidative Stress Models: Investigated for roles in redox homeostasis under stress conditions (e.g., methyl viologen exposure) .
Comparative Analysis with Homologs
The Haemophilus influenzae NapH homolog (UniProt ID: P44653) shares 65% sequence identity with E. coli NapH but exhibits distinct biochemical properties, such as altered solubility profiles .
| Feature | E. coli NapH | H. influenzae NapH |
|---|---|---|
| Sequence Identity | 100% | 65% |
| Solubility | Partial in cytoplasm | Mostly insoluble |
| Tag | His tag | His tag |
Challenges and Future Directions
Product Specs
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Target Background
KEGG: ecj:JW2192
STRING: 316385.ECDH10B_2361
