X-ray Structure of Human Class IV ςς Alcohol Dehydrogenase

Xie, Peiguang; Parsons, Stephen H.; Speckhard, David C.; Bosron, William F.; Hurley, Thomas D.

doi:10.1074/jbc.272.30.18558

Cited by 79 publications

(41 citation statements)

References 33 publications

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“…51). 4-MP interacts with positions 94 (54) and 142 (55). Class I ADHs have Phe 94 and Leu 142 , whereas the class III enzymes have Tyr 94 and Met 142 .…”

Section: Figmentioning

confidence: 99%

Two Zebrafish Alcohol Dehydrogenases Share Common Ancestry with Mammalian Class I, II, IV, and V Alcohol Dehydrogenase Genes but Have Distinct Functional Characteristics

Reimers

Hahn

Tanguay

2004

Journal of Biological Chemistry

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Ethanol is teratogenic to many vertebrates. We are utilizing zebrafish as a model system to determine whether there is an association between ethanol metabolism and ethanol-mediated developmental toxicity. Here we report the isolation and characterization of two cDNAs encoding zebrafish alcohol dehydrogenases (ADHs). Phylogenetic analysis of these zebrafish ADHs indicates that they share a common ancestor with mammalian class I, II, IV, and V ADHs. The genes encoding these zebrafish ADHs have been named Adh8a and Adh8b by the nomenclature committee. Both genes were genetically mapped to chromosome 13. The 1450-bp Adh8a is 82, 73, 72, and 72% similar at the amino acid level to the Baltic cod ADH8 (previously named ADH1), the human ADH1B2, the mouse ADH1, and the rat ADH1, respectively. Also, the 1484-bp Adh8b is 77, 68, 67, and 66% similar at the amino acid level to the Baltic cod ADH8, the human ADH1B2, the mouse ADH1, and the rat ADH1, respectively. ADH8A and ADH8B share 86% amino acid similarity. To characterize the functional properties of ADH8A and ADH8B, recombinant proteins were purified from SF-9 insect cells. Kinetic studies demonstrate that ADH8A metabolizes ethanol, with a V max of 13.4 nmol/min/mg protein, whereas ADH8B does not metabolize ethanol. The ADH8A K m for ethanol as a substrate is 0.7 mM. 4-Methyl pyrazole, a classical competitive inhibitor of class I ADH, failed to inhibit ADH8A. ADH8B has the capacity to efficiently biotransform longer chain primary alcohols (>5 carbons) and S-hydroxymethlyglutathione, whereas ADH8A does not efficiently metabolize these substrates. Finally, mRNA expression studies indicate that both ADH8A and ADH8B mRNA are expressed during early development and in the adult brain, fin, gill, heart, kidney, muscle, and liver. Together these results indicate that class Ilike ADH is conserved in zebrafish, albeit with mixed functional properties.Fetal alcohol syndrome in children born to women who consumed alcohol during pregnancy was first described by Jones and colleagues in 1973 (1). Fetal alcohol syndrome is characterized by a delay in development, cardiac abnormalities (1), central nervous abnormalities, abnormal craniofacial features, and intellectual delays (1, 2). The teratogenic properties of ethanol have been firmly established; however, the underlying mechanism(s) of toxicity remain unclear. Two molecular mechanisms have been proposed: direct ethanol effects and the indirect effects associated with acetaldehyde formation and oxidative stress (3). The ability of ethanol to cause developmental anomalies has been demonstrated in mice, rats, Drosophila melanogaster, and chickens (4 -7). Zebrafish are well suited for genetic studies (reviewed in Ref. 8). Zebrafish embryos exposed to ethanol display craniofacial abnormalities, cardiac and structural malformations, and development delay (9 -11). Visual function was affected in embryos exposed to 1.5% ethanol during development (12). Recent studies indicate that three adult strains of zebrafish display different behavior...

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“…51). 4-MP interacts with positions 94 (54) and 142 (55). Class I ADHs have Phe 94 and Leu 142 , whereas the class III enzymes have Tyr 94 and Met 142 .…”

Section: Figmentioning

confidence: 99%

Two Zebrafish Alcohol Dehydrogenases Share Common Ancestry with Mammalian Class I, II, IV, and V Alcohol Dehydrogenase Genes but Have Distinct Functional Characteristics

Reimers

Hahn

Tanguay

2004

Journal of Biological Chemistry

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“…No solution has yet been obtained by molecular replacement with the programs AMoRe (Navaza, 1994) and BEAST (Read, 2001) using search models derived from human ADH1B1 (Hurley et al, 1991), human ADH4 (Xie et al, 1997), horse ADH1 (Adolph et al, 2000) and ketose reductase (Ban®eld et al, 2001) and from combinations of the N-terminal catalytic and coenzyme-binding domains of these structures. All ADH molecules used as search models exhibit sequence identities with ScADHVI of below 25% which, together Crystals of (a) ScADHVI and (b) ADH8.…”

Section: Resultsmentioning

confidence: 99%

“…No crystallographic structure has been reported for any member of the CAD family. The closest known structures to ADHVI (25% identity) are of a NADP(H)-dependent ketose reductase from the white¯y Bemisia argentifolii (Ban®eld et al, 2001) and human ADH4 (Xie et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

“…Several crystallographic structures of vertebrate ADHs are known: horse ADH1 (Eklund et al, 1976), human ADH1B1, ADH1B2 and ADH1B3 , human ADH1A and ADH1C2 (Niederhut et al, 2001), cod ADH1 (Ramaswamy et al, 1996), mouse ADH2 (Svensson et al, 2000), human ADH3 (Yang et al, 1997) and human ADH4 (Xie et al, 1997). All of them correspond to NAD(H)-dependent enzymes and exhibit less than 57% identity with the present amphibian ADH8.…”

Section: Introductionmentioning

confidence: 99%

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Crystallization and preliminary X-ray analysis of NADP(H)-dependent alcohol dehydrogenases fromSaccharomyces cerevisiaeandRana perezi

Valencia

Rosell

Larroy

et al. 2003

Acta Crystallogr D Biol Cryst

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Different crystal forms diffracting to high resolution have been obtained for two NADP(H)-dependent alcohol dehydrogenases, members of the medium-chain dehydrogenase/reductase superfamily: ScADHVI from Saccharomyces cerevisiae and ADH8 from Rana perezi. ScADHVI is a broad-speci®city enzyme, with a sequence identity lower than 25% with respect to all other ADHs of known structure. The best crystals of ScADHVI diffracted beyond 2.8 A Ê resolution and belonged to the trigonal space group P3 1 21 (or to its enantiomorph P3 2 21), with unit-cell parameters a = b = 102.2, c = 149.7 A Ê , = 120 . These crystals were produced by the hangingdrop vapour-diffusion method using ammonium sulfate as precipitant. Packing considerations together with the self-rotation function and the native Patterson map seem to indicate the presence of only one subunit per asymmetric unit, with a volume solvent content of about 80%. ADH8 from R. perezi is the only NADP(H)-dependent ADH from vertebrates characterized to date. Crystals of ADH8 obtained both in the absence and in the presence of NADP + using polyethylene glycol and lithium sulfate as precipitants diffracted to 2.2 and 1.8 A Ê , respectively, using synchrotron radiation. These crystals were isomorphous, space group C2, with approximate unitcell parameters a = 122, b = 79, c = 91 A Ê , = 113 and contain one dimer per asymmetric unit, with a volume solvent content of about 50%.

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“…ADH class I and class IV have an activity in the first step of RA synthesis [16,17]. Crystal structures of ADH class I and class IV revealed a conserved active site that can accommodate long chain alcohols such as retinol [18][19][20]. ADH class I is expressed at high levels in many retinoid target tissues of embryos [15,21] and in adult epithelia, such as intestine, kidney, adrenal gland, testis, epididymis, uterus and ovary [15,[22][23][24][25].…”

Section: Discussionmentioning

confidence: 99%

Evidence for the Expression of Alcohol Dehydrogenase Class I Gene in Rat Uterus and Its Up-regulation by Progesterone

et al. 2008

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Abstract. The endometrium is one of the target tissues of the ovarian steroid hormones, estrogen and progesterone. In order to elucidate the mechanism of gene regulation in the endometrium, suppressive subtraction hybridization was performed to isolate the candidate genes controlled by progesterone in rat uterus. Alcohol dehydrogenase (ADH) class I gene was one of the candidate genes. Here we investigated the expression and regulation of ADH class I gene in rat uterus. The mRNA of ADH class I was detected in uterus by RT-PCR using specific primers. Using specific probe for ADH class I, in situ hybridization was performed to investigate localization in rat uterus. Positive signals were detected in the endometrial stromal cells of rat uterus by in situ hybridization and were not detected in endometrial epithelial cells and myometrium in rat uterus. Ovariectomized rats were treated with 17-β estradiol and progesterone and the uteri of these rats were used for Northern blot analysis and assay of the ADH activity. Northern blot analysis revealed that the expression of ADH class I mRNA in rat uteri was up-regulated approximately two-fold after progesterone treatment, but not estrogen. Likewise, ADH activity was approximately two-fold higher in progesterone-treated rat uteri compared with controls. This study demonstrated that ADH class I gene is progesterone-responsive in the uterus. This implies that progesterone might be involved with retinoic acid synthesis in the uterus, since ADH is the key enzyme for retinoic acid synthesis.

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X-ray Structure of Human Class IV ςς Alcohol Dehydrogenase

Abstract: The structural determinants of substrate recognition in the human class IV, or , alcohol dehydrogenase (ADH) isoenzyme were examined through x-ray crystallography and site-directed mutagenesis. The crystal structure of ADH complexed with NAD

Cited by 79 publications

References 33 publications

Two Zebrafish Alcohol Dehydrogenases Share Common Ancestry with Mammalian Class I, II, IV, and V Alcohol Dehydrogenase Genes but Have Distinct Functional Characteristics

Two Zebrafish Alcohol Dehydrogenases Share Common Ancestry with Mammalian Class I, II, IV, and V Alcohol Dehydrogenase Genes but Have Distinct Functional Characteristics

Crystallization and preliminary X-ray analysis of NADP(H)-dependent alcohol dehydrogenases fromSaccharomyces cerevisiaeandRana perezi

Evidence for the Expression of Alcohol Dehydrogenase Class I Gene in Rat Uterus and Its Up-regulation by Progesterone

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