ArticleProtein science : a publication of the Protein Society2004
Removal of N-terminal methionine from recombinant proteins by engineered E. coli methionine aminopeptidase.
Article in Protein science : a publication of the Protein Society, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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30 citing papers in PubMed, 79 citations in OpenAlex.
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- Strategies to Enhance Periplasmic Recombinant Protein Production Yields inFrontiers in bioengineering and biotechnology · 2021Review
- Development of Native MS Capabilities on an Extended Mass Range Q-TOF MS.International journal of mass spectrometry · 2020Article
- Analysis of Molecular Heterogeneity in Therapeutic IFNα2b from Different Manufacturers by LC/Q-TOF.Molecules (Basel, Switzerland) · 2020Article
- Heterologous expression of biologically active Mambalgin-1 peptide as a new potential anticancer, using a PVX-based viral vector inPlant cell, tissue and organ culture · 2020Article
- Segments in the Amyloid Core that Distinguish Hamster from Mouse Prion Fibrils.Neurochemical research · 2019Article
- High throughput discovery of protein variants using proteomics informed by transcriptomics.Nucleic acids research · 2018Article
- Refining the Ambush Hypothesis: Evidence That GC- and AT-Rich Bacteria Employ Different Frameshift Defence Strategies.Genome biology and evolution · 2018Article
- Engineering posttranslational proofreading to discriminate nonstandard amino acids.Proceedings of the National Academy of Sciences of the United States of America · 2018Article
- Biochemical and in silico evaluation of recombinant E. coli aminopeptidase and in vitro processed human interferon α-2b.Turkish journal of biology = Turk biyoloji dergisi · 2018Article
- Adenine Enrichment at the Fourth CDS Residue in Bacterial Genes Is Consistent with Error Proofing for +1 Frameshifts.Molecular biology and evolution · 2017Article
- Targeting the N terminus for site-selective protein modification.Nature chemical biology · 2017Review
- N-Terminal Methionine Processing.Current protocols in protein science · 2017Article
- A brush-polymer conjugate of exendin-4 reduces blood glucose for up to five days and eliminates poly(ethylene glycol) antigenicity.Nature biomedical engineering · 2016Article
- Overview of the purification of recombinant proteins.Current protocols in protein science · 2015Review
- Motif-directed redesign of enzyme specificity.Protein science : a publication of the Protein Society · 2014Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The removal of N-terminal translation initiator Met by methionine aminopeptidase (MetAP) is often crucial for the function and stability of proteins. On the basis of crystal structure and sequence alignment of MetAPs, we have engineered Escherichia coli MetAP by the mutation of three residues, Y168G, M206T, Q233G, in the substrate-binding pocket. Our engineered MetAPs are able to remove the Met from bulky or acidic penultimate residues, such as Met, His, Asp, Asn, Glu, Gln, Leu, Ile, Tyr, and Trp, as well as from small residues. The penultimate residue, the second residue after Met, was further removed if the antepenultimate residue, the third residue after Met, was small. By the coexpression of engineered MetAP in E. coli through the same or a separate vector, we have successfully produced recombinant proteins possessing an innate N terminus, such as onconase, an antitumor ribonuclease from the frog Rana pipiens. The N-terminal pyroglutamate of recombinant onconase is critical for its structural integrity, catalytic activity, and cyto-toxicity. On the basis of N-terminal sequence information in the protein database, 85%-90% of recombinant proteins should be produced in authentic form by our engineered MetAPs.
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