Incorporation of an artificial receptor into a native protein: New strategy for the design of semisynthetic enzymes with allosteric properties

Itaru Hamachi, Tsuyoshi Nagase, Yusuke Tajiri, Seiji Shinkai

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The sugar-facilitated structure and enzymatic activity change of engineered myoglobins bearing a phenylboronic acid moiety, which were semisynthesized by a cofactor reconstitution method, were studied by the denaturation experiment, spectrophotometric titration of the pK(a) shift of the axial H2O, circular dichloism (CD), and the kinetics of the myoglobin-catalyzed-aniline hydroxylation reaction. Both boronophenylalanine-appended myoglobin [Mb(m-Bphe)2] and phenylboronic acid-appended myoglobin [Mb(PhBOH)2] were stabilized by approximately 2 kcal/mol upon complexation with D-fructose. CD spectral changes and the sugar-induced pK(a) shift suggested that the microenvironment of the active site of these myoglobins was re-formed from a partially disturbed state to that comparable to the native state upon D-fructose binding. The correlation of pK(a) with k(cat) (for the aniline hydroxylase activity) and the ΔG(D)(H2O)-k(cat) profile showed that these structural changes of Mb-(m-Bphe)2 and Mb(PhBOH)2 were closely related to their sugar-enhanced aniline hydroxylase activity. Thus, the results established that an incorporation of the artificial receptor molecule can be a valid methodology for the design of stimuli-responsive semiartificial enzymes.

Original languageEnglish
Pages (from-to)862-868
Number of pages7
JournalBioconjugate Chemistry
Volume8
Issue number6
DOIs
Publication statusPublished - Nov 1 1997

Fingerprint

Artificial Receptors
Myoglobin
Aniline
Sugars
Fructose
Enzymes
Proteins
Aniline Hydroxylase
Bearings (structural)
Hydroxylation
Denaturation
Acids
Complexation
Titration
Molecules
Kinetics
Catalytic Domain
Experiments

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

Cite this

Incorporation of an artificial receptor into a native protein : New strategy for the design of semisynthetic enzymes with allosteric properties. / Hamachi, Itaru; Nagase, Tsuyoshi; Tajiri, Yusuke; Shinkai, Seiji.

In: Bioconjugate Chemistry, Vol. 8, No. 6, 01.11.1997, p. 862-868.

Research output: Contribution to journalArticle

Hamachi, Itaru ; Nagase, Tsuyoshi ; Tajiri, Yusuke ; Shinkai, Seiji. / Incorporation of an artificial receptor into a native protein : New strategy for the design of semisynthetic enzymes with allosteric properties. In: Bioconjugate Chemistry. 1997 ; Vol. 8, No. 6. pp. 862-868.
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