Enzymatic characteristics of retinal dehydrogenase type I expressed in Escherichia coli

Peter Penzes; Xianshu Wang; Joseph L. Napoli

doi:10.1016/S0167-4838(97)00102-7

Enzymatic characteristics of retinal dehydrogenase type I expressed in Escherichia coli

Peter Penzes, Xianshu Wang, Joseph L. Napoli^*

^*Corresponding author for this work

Neuroscience

Research output: Contribution to journal › Article › peer-review

38 Scopus citations

Abstract

We expressed RalDH(I) in Escherichia coli and have shown that it functions in vitro with the complex CRBP-retinal (cellular retinol-binding protein) as substrate, either generated in situ from the complex of CRBP-retinol and microsomal retinol dehydrogenases or provided directly as CRBP-retinal. Recombinant RalDH(I) had kinetic constants with CRBP-retinal of: Hill coefficient 1.8; K_0.5 0.8 μM; and V(m) 1.5 nmol/min/mg of protein at 25°C. Apo-CRBP inhibited the reaction with CRBP-rerinal with an IC₅₀ of 1.4 μM. Citral inhibited RalDH(I) with an IC₅₀ of ~ 1 μM compared to an IC₅₀ of ~ 12 for RalDH(II), but did not serve as substrate for RalDH(I). RalDH(I) did not catalyze efficiently the dehydrogenation of acetaldehyde, but showed higher V(max)/K(m) values for hexanal, octanal, decanal and benzaldehyde than for either propanal or retinal. These data extend the characterization of RalDH(I), show that apo-CRBP competes with holo-CRBP as substrate for RalDH(I), and expand insight into the pathways of retinoic acid biogenesis from the most abundant substratcs in vivo, retinoid-liganded CRBP.

Original language	English (US)
Pages (from-to)	175-181
Number of pages	7
Journal	Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
Volume	1342
Issue number	2
DOIs	https://doi.org/10.1016/S0167-4838(97)00102-7
State	Published - Oct 17 1997

Funding

We are grateful for critical input from Edward G. Niles throughout this project. This work was supported by National Institutes of Health Grant AG13566.

Keywords

Aldehyde dehydrogenase
Retinal
Retinal dehydrogenase
Retinoic acid
Retinol

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology

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10.1016/S0167-4838(97)00102-7

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title = "Enzymatic characteristics of retinal dehydrogenase type I expressed in Escherichia coli",

abstract = "We expressed RalDH(I) in Escherichia coli and have shown that it functions in vitro with the complex CRBP-retinal (cellular retinol-binding protein) as substrate, either generated in situ from the complex of CRBP-retinol and microsomal retinol dehydrogenases or provided directly as CRBP-retinal. Recombinant RalDH(I) had kinetic constants with CRBP-retinal of: Hill coefficient 1.8; K0.5 0.8 μM; and V(m) 1.5 nmol/min/mg of protein at 25°C. Apo-CRBP inhibited the reaction with CRBP-rerinal with an IC50 of 1.4 μM. Citral inhibited RalDH(I) with an IC50 of ~ 1 μM compared to an IC50 of ~ 12 for RalDH(II), but did not serve as substrate for RalDH(I). RalDH(I) did not catalyze efficiently the dehydrogenation of acetaldehyde, but showed higher V(max)/K(m) values for hexanal, octanal, decanal and benzaldehyde than for either propanal or retinal. These data extend the characterization of RalDH(I), show that apo-CRBP competes with holo-CRBP as substrate for RalDH(I), and expand insight into the pathways of retinoic acid biogenesis from the most abundant substratcs in vivo, retinoid-liganded CRBP.",

keywords = "Aldehyde dehydrogenase, Retinal, Retinal dehydrogenase, Retinoic acid, Retinol",

author = "Peter Penzes and Xianshu Wang and Napoli, {Joseph L.}",

note = "Funding Information: We are grateful for critical input from Edward G. Niles throughout this project. This work was supported by National Institutes of Health Grant AG13566.",

year = "1997",

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doi = "10.1016/S0167-4838(97)00102-7",

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T1 - Enzymatic characteristics of retinal dehydrogenase type I expressed in Escherichia coli

AU - Penzes, Peter

AU - Wang, Xianshu

AU - Napoli, Joseph L.

N1 - Funding Information: We are grateful for critical input from Edward G. Niles throughout this project. This work was supported by National Institutes of Health Grant AG13566.

PY - 1997/10/17

Y1 - 1997/10/17

N2 - We expressed RalDH(I) in Escherichia coli and have shown that it functions in vitro with the complex CRBP-retinal (cellular retinol-binding protein) as substrate, either generated in situ from the complex of CRBP-retinol and microsomal retinol dehydrogenases or provided directly as CRBP-retinal. Recombinant RalDH(I) had kinetic constants with CRBP-retinal of: Hill coefficient 1.8; K0.5 0.8 μM; and V(m) 1.5 nmol/min/mg of protein at 25°C. Apo-CRBP inhibited the reaction with CRBP-rerinal with an IC50 of 1.4 μM. Citral inhibited RalDH(I) with an IC50 of ~ 1 μM compared to an IC50 of ~ 12 for RalDH(II), but did not serve as substrate for RalDH(I). RalDH(I) did not catalyze efficiently the dehydrogenation of acetaldehyde, but showed higher V(max)/K(m) values for hexanal, octanal, decanal and benzaldehyde than for either propanal or retinal. These data extend the characterization of RalDH(I), show that apo-CRBP competes with holo-CRBP as substrate for RalDH(I), and expand insight into the pathways of retinoic acid biogenesis from the most abundant substratcs in vivo, retinoid-liganded CRBP.

AB - We expressed RalDH(I) in Escherichia coli and have shown that it functions in vitro with the complex CRBP-retinal (cellular retinol-binding protein) as substrate, either generated in situ from the complex of CRBP-retinol and microsomal retinol dehydrogenases or provided directly as CRBP-retinal. Recombinant RalDH(I) had kinetic constants with CRBP-retinal of: Hill coefficient 1.8; K0.5 0.8 μM; and V(m) 1.5 nmol/min/mg of protein at 25°C. Apo-CRBP inhibited the reaction with CRBP-rerinal with an IC50 of 1.4 μM. Citral inhibited RalDH(I) with an IC50 of ~ 1 μM compared to an IC50 of ~ 12 for RalDH(II), but did not serve as substrate for RalDH(I). RalDH(I) did not catalyze efficiently the dehydrogenation of acetaldehyde, but showed higher V(max)/K(m) values for hexanal, octanal, decanal and benzaldehyde than for either propanal or retinal. These data extend the characterization of RalDH(I), show that apo-CRBP competes with holo-CRBP as substrate for RalDH(I), and expand insight into the pathways of retinoic acid biogenesis from the most abundant substratcs in vivo, retinoid-liganded CRBP.

KW - Aldehyde dehydrogenase

KW - Retinal

KW - Retinal dehydrogenase

KW - Retinoic acid

KW - Retinol

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JO - Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology

JF - Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology

IS - 2

ER -

Enzymatic characteristics of retinal dehydrogenase type I expressed in Escherichia coli

Abstract

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