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Substrate specificities of deuterolysin from Aspergillus oryzae and electron paramagnetic resonance measurement of cobalt-substituted deuterolysin
- Source :
- Bioscience, biotechnology, and biochemistry. 67(2)
- Publication Year :
- 2003
-
Abstract
- The substrate specificities of deuterolysin, a 19-kDa zinc-protease (EC 3.4.24.39) from Aspergillus oryzae, were investigated at pH 9.0 with various fluorogenic acyl-peptide-4-methylcoumaryl-7-amides (peptide-MCAs). N-Butoxycarbonyl-Arg-Val-Arg-Arg-MCA was the best substrate for deuterolysin. We therefore measured its kinetic parameters. Deuterolysin had high activity toward the peptide bonds next to pairs of basic residues in calf thymus histone H4. The specificity of cobalt-substituted deuterolysin (Co-deuterolysin) for peptide-MCAs was similar to that of native deuterolysin. The CD spectrum of Co-deuterolysin was similar to that of the native deuterolysin. The metal coordination sphere of Co-deuterolysin was analyzed by Q-band (33.9570 GHz) electron paramagnetic resonance (EPR) spectroscopy. Using computer simulation of EPR, we found the g principal values to be g(xx) = 5.20, g(yy) = 4.75, and g(zz) = 2.24; the metal center was a divalent cobalt ion in a high spin state.
- Subjects :
- Coordination sphere
Hot Temperature
Aspergillus oryzae
Molecular Sequence Data
chemistry.chemical_element
Peptide
Thymus Gland
Applied Microbiology and Biotechnology
Biochemistry
Analytical Chemistry
law.invention
Divalent
Substrate Specificity
Histones
law
Enzyme Stability
Peptide bond
Organic chemistry
Animals
Computer Simulation
Amino Acid Sequence
Electron paramagnetic resonance
Molecular Biology
chemistry.chemical_classification
Binding Sites
biology
Amino Acids, Basic
Circular Dichroism
Organic Chemistry
Electron Spin Resonance Spectroscopy
Substrate (chemistry)
Metalloendopeptidases
General Medicine
Cobalt
biology.organism_classification
Amides
Crystallography
Kinetics
Zinc
chemistry
Cattle
Biotechnology
Subjects
Details
- ISSN :
- 09168451
- Volume :
- 67
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Bioscience, biotechnology, and biochemistry
- Accession number :
- edsair.doi.dedup.....de094f687be9b6eeb9ebc7fef09bf2b9