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Expression, characterization, and reaction of recombinant monkey metallothionein-1 and its C33M mutant
- Source :
- Journal of protein chemistry. 21(3)
- Publication Year :
- 2002
-
Abstract
- After we modified the protocol of purification, monkey metallothionein-1 (mkMT-1) and its mutant at position 33 (C33M mutant) were efficiently expressed and purified by using the glutathione-S-transferase fusion protein system. The protein yield has been considerably improved (8 mg/L culture for mkMT-1 and 10 mg/L culture for C33M mutant). The recombinant MT-1 and C33M mutant were characterized by ESI-MS, UV, and CD spectra. The reactions of MI-1 and C33M mutant with 5,5'-dithiobis(2-nitrobenzoic acid) and EDTA also have been carefully studied. The pH titration of MT-1 and C33M mutant has been studied by UV and CD spectra. The mutation of cysteine-to-methionine at position 33 mostly maintains the alpha-domain structure similar to that in wild-type mkMT-1, but the C33M mutant has significant loss of stability and cooperative properties of the domain.
- Subjects :
- Recombinant Fusion Proteins
Mutant
Molecular Sequence Data
Mutagenesis (molecular biology technique)
Dithionitrobenzoic Acid
Biology
medicine.disease_cause
Biochemistry
law.invention
law
medicine
Escherichia coli
Metallothionein
Bioorganic chemistry
Animals
Amino Acid Sequence
Edetic Acid
Glutathione Transferase
Mutation
Circular Dichroism
Titrimetry
Haplorhini
Hydrogen-Ion Concentration
Fusion protein
Molecular biology
Molecular Weight
Kinetics
Recombinant DNA
Mutagenesis, Site-Directed
Titration
Electrophoresis, Polyacrylamide Gel
Spectrophotometry, Ultraviolet
Cadmium
Subjects
Details
- ISSN :
- 02778033
- Volume :
- 21
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Journal of protein chemistry
- Accession number :
- edsair.doi.dedup.....1886e284e5573b9bc23d162b561bd8d2