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Volumn 8, Issue 4, 1998, Pages 459-465

The design of protein-based catalysts using semisynthetic methods

Author keywords

[No Author keywords available]

Indexed keywords

AMINO ACID; AMINOTRANSFERASE; PYRIDOXAL; PYRIDOXAMINE;

EID: 0032143901     PISSN: 0959440X     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0959-440X(98)80123-1     Document Type: Article
Times cited : (22)

References (53)
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    • of outstanding interest. Using a series of four mutants of the E. coli RNA polymerase α subunit that each contain a single cysteine residue, four EDTA conjugates were prepared. It was necessary to prepare the constructs under denaturing conditions in order to obtain efficient conjugation. These modified proteins were used in affinity-cleavage experiments to show that the N-terminal assembly domain of one subunit is proximal to the linker region between the N- and C-terminal domains of the other subunit. In contrast to previous results with other EDTA-based conjugates, cleavage may occur in these examples via oxidative processes.
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    • of special interest. ALBP - PX, an adipocyte lipid-binding protein conjugate that positions a disulfide-bound pyridoxamine cofactor within a sequestered protein cavity, reductively aminates a number of alkyl, aryl and sidechain functionalized α-keto acids to α-amino acids with enantioselectivities as high as 94% ee.
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    • Structural characterization of two synthetic catalysts based on adipocyte lipid binding protein
    • of outstanding interest. The structures, determined by X-ray diffraction analysis, of two conjugates based on the adipocyte lipid binding protein are described. The X-ray models show the presence of the phenanthroline and pyridoxamine moieties within the protein cavity. They are useful for examining the origin of the chiral selectivities seen in the reactions catalyzed by these constructs. In both cases, these covalent adducts reduce the volume of the internal cavity and sterically limit substrate interactions as well as solvent access to potential intermediates in the reaction pathway.
    • Ory JJ, Mazhary A, Kuang H, Davies RR, Distelano MD, Banaszak LJ. Structural characterization of two synthetic catalysts based on adipocyte lipid binding protein. of outstanding interest Protein Eng. 11:1998;253-261 The structures, determined by X-ray diffraction analysis, of two conjugates based on the adipocyte lipid binding protein are described. The X-ray models show the presence of the phenanthroline and pyridoxamine moieties within the protein cavity. They are useful for examining the origin of the chiral selectivities seen in the reactions catalyzed by these constructs. In both cases, these covalent adducts reduce the volume of the internal cavity and sterically limit substrate interactions as well as solvent access to potential intermediates in the reaction pathway.
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    • Modulation of the rate, enantioselectivity and substrate specificity of semisynthetic transaminases based on lipid binding proteins using site directed mutagenesis
    • of special interest. A series of constructs in which the position of the pyridoxamine attachment within the intestinal fatty acid binding protein (IFABP) cavity was varied by site-directed mutagenesis are described. The IFABP-PX60 conjugate reacts at least 9.4-fold more rapidly than the original ALBP-PX conjugate, whereas IFABP-PX72 inverts the enantioselectivity of the reactions (compared to ALBP-PX) and IFABP-PX104 displays very selective substrate specificity. These results indicate that site-directed mutagenesis can be used to tune the rate, enantioselectivity and substrate specificity of semisynthetic transaminases that are based on fatty acid-binding proteins.
    • Kuang H, Davies RR, Distefano MD. Modulation of the rate, enantioselectivity and substrate specificity of semisynthetic transaminases based on lipid binding proteins using site directed mutagenesis. of special interest Bioorg Med Chem Lett. 7:1997;2055-2060 A series of constructs in which the position of the pyridoxamine attachment within the intestinal fatty acid binding protein (IFABP) cavity was varied by site-directed mutagenesis are described. The IFABP-PX60 conjugate reacts at least 9.4-fold more rapidly than the original ALBP-PX conjugate, whereas IFABP-PX72 inverts the enantioselectivity of the reactions (compared to ALBP-PX) and IFABP-PX104 displays very selective substrate specificity. These results indicate that site-directed mutagenesis can be used to tune the rate, enantioselectivity and substrate specificity of semisynthetic transaminases that are based on fatty acid-binding proteins.
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    • Catalytic enantioselective reductive amination in a host-guest system based on a protein cavity
    • of outstanding interest. The properties of IFABP-PX60, a catalyst constructed by the covalent attachment of a pyridoxamine cofactor to a cysteine residue within the cavity of a mutant form of the intestinal fatty acid-binding protein (IFABP), are reported. Under single turnover conditions, IFABP-PX60 converts α-keto glutarate to glutamic acid 62-fold faster than free pyridoxamine. This conjugate catalytically performs this reaction with an efficiency at least 200-fold greater than the free cofactor; as many as 50 turnovers with an enantiomeric purity of 95% ee have been obtained. These results indicate that a host-guest system based on a fatty acid-binding protein can be used to generate a construct that mimics three key features of enzymatic catalysis: selectivity, rate enhancement and turnover.
    • Kuang H, Distefano MD. Catalytic enantioselective reductive amination in a host-guest system based on a protein cavity. of outstanding interest J Am Chem Soc. 120:1998;1072-1073 The properties of IFABP-PX60, a catalyst constructed by the covalent attachment of a pyridoxamine cofactor to a cysteine residue within the cavity of a mutant form of the intestinal fatty acid-binding protein (IFABP), are reported. Under single turnover conditions, IFABP-PX60 converts α-keto glutarate to glutamic acid 62-fold faster than free pyridoxamine. This conjugate catalytically performs this reaction with an efficiency at least 200-fold greater than the free cofactor; as many as 50 turnovers with an enantiomeric purity of 95% ee have been obtained. These results indicate that a host-guest system based on a fatty acid-binding protein can be used to generate a construct that mimics three key features of enzymatic catalysis: selectivity, rate enhancement and turnover.
    • (1998) J Am Chem Soc , vol.120 , pp. 1072-1073
    • Kuang, H.1    Distefano, M.D.2
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    • Effects of metal ions on the rates and enantioselectivities of reactions catalyzed by a series of semisynthetic transaminases created by site directed mutagenesis
    • of special interest. The chemistry performed by intestinal fatty acid-binding protein (IFABP) conjugates under catalytic conditions and the effect on the reaction rate and enantioselectivity of added metal ions are both described. It appears that metal ions can be used to increase the rate of reactions catalyzed by semisynthetic transaminases, although the addition of metal ions can also retard the reaction rate. Furthermore, the presence of metal ions decreases reaction enantioselectivity.
    • Qi D, Kuang H, Distefano MD. Effects of metal ions on the rates and enantioselectivities of reactions catalyzed by a series of semisynthetic transaminases created by site directed mutagenesis. of special interest Bioorg Med Chem Lett. 8:1998;875-880 The chemistry performed by intestinal fatty acid-binding protein (IFABP) conjugates under catalytic conditions and the effect on the reaction rate and enantioselectivity of added metal ions are both described. It appears that metal ions can be used to increase the rate of reactions catalyzed by semisynthetic transaminases, although the addition of metal ions can also retard the reaction rate. Furthermore, the presence of metal ions decreases reaction enantioselectivity.
    • (1998) Bioorg Med Chem Lett , vol.8 , pp. 875-880
    • Qi, D.1    Kuang, H.2    Distefano, M.D.3
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    • A semisynthetic metalloenzyme based on a protein cavity that catalyzes the enantioselective hydrolysis of ester and amide substrates
    • 4-fold above the background rate. The kinetics of this amide hydrolysis reaction fit the Michaelis-Menten relationship that is characteristic of enzymatic processes.
    • 4-fold above the background rate. The kinetics of this amide hydrolysis reaction fit the Michaelis-Menten relationship that is characteristic of enzymatic processes.
    • (1997) J Am Chem Soc , vol.119 , pp. 11643-11652
    • Davies, R.R.1    Aidistefano, M.D.2
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    • MOLSCRIPT; A program to produce both detailed and schematic plots of protein structure
    • Kraulis PJ. MOLSCRIPT; a program to produce both detailed and schematic plots of protein structure. J Appl Crystallogr. 24:1991;946-950.
    • (1991) J Appl Crystallogr , vol.24 , pp. 946-950
    • Kraulis, P.J.1


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.