Crystallization and Preliminary X-ray Diffraction Studies on Cytosolic (Class 1) Aldehyde Dehydrogenase from Sheep Liver

Baker, Heather M.; Brown, Rosemary L.; Dobbs, Aaron J.; Blackwell, Leonard F.; Buckley, Paul D.; Hardman, Michael J.; Hill, J. P.; Kitson, Kathryn E.; Kitson, Trevor M.; Baker, Edward N.

doi:10.1006/jmbi.1994.1494

Cited by 6 publications

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“…The purest fractions were then pooled, dialysed against 50 mM Na/K phosphate, pH 7.4, 1 mM DTT and concentrated to 10 mg/ml protein using microconcentrators. Crystals were grown using the hanging-drop vapour diffusion technique as described previously [48]. The protein concentration was 10 mg/ml with 1 mM NAD + , and the reservoir solution contained 170 mM MgCl 2 , 100 mM Bis-Tris-propane (pH 6-7) and 6-7% w/v monomethylether polyethylene glycol (PEG) 5000.…”

Section: Isolation Purification and Crystallisation Of Sheep Liver Aldh1mentioning

confidence: 99%

Sheep liver cytosolic aldehyde dehydrogenase: the structure reveals the basis for the retinal specificity of class 1 aldehyde dehydrogenases

et al. 1998

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. The submicromolar Km of ALDH1 for all-trans retinal, and its 600-fold enhanced affinity for retinal compared to acetaldehyde, are explained by the size and shape of the substrate entrance tunnel in ALDH1. All-trans retinal fits into the active-site pocket of ALDH1, but not into the pocket of ALDH2. Two helices and one surface loop that line the tunnel are likely to have a key role in defining substrate specificity in the wider ALDH family. The relative sizes of the tunnels also suggest why the bulky alcohol aversive drug disulfiram reacts more rapidly with ALDH1 than ALDH2. The disorder of Glu268 and the observation that NAD+ binds in two distinct modes indicate that flexibility is a key facet of the enzyme reaction mechanism.

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Section: Isolation Purification and Crystallisation Of Sheep Liver Aldh1mentioning

confidence: 99%

Sheep liver cytosolic aldehyde dehydrogenase: the structure reveals the basis for the retinal specificity of class 1 aldehyde dehydrogenases

et al. 1998

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Crystallization and preliminary X‐ray analysis of aldehyde dehydrogenase from Vibrio harveyi

et al. 1996

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Aldehyde dehydrogenase from Vibrio harveyi catalyzes the oxidation of long-chain aliphatic aldehydes to acids. The enzyme is unique among the family of aldehyde dehydrogenases in that it exhibits much higher specificity for the cofactor NADP+ than for NAD+. The sequence of this form of the enzyme varies significantly from the NAD' dependent forms, suggesting differences in the three-dimensional structure that may be correlated to cofactor specificity. Crystals of the enzyme have been grown both in the presence and absence of NADP' using the hanging drop vapor diffusion technique. In order to improve crystal size and quality, iterative seeding techniques were employed. The crystals belong to space group P2,, with unit cell dimensions a = 79.4 A, b = 131 . I A, c = 92.2 A, and p = 92.4". Freezing the crystal to IOOK has enabled a complete set of data to be collected using a rotating anode source (A = 1.541 8 A). The crystals diffract to a minimum d-spacing of 2.6 A resolution. Based on density calculations, two homodimers of molecular weight 110 kDa are estimated to be present in the asymmetric unit. Self-rotation functions show the presence of 3 noncrystallographic twofold symmetry axes.Keywords: aldehyde dehydrogenase; crystallization; X-ray diffraction NADP' specificity Aldehyde dehydrogenases constitute a large family of NAD(P)'-dependent enzymes found in both prokaryotes and eukaryotes. The enzyme catalyzes the dehydrogenation of aldehydes via the formation of a thiohemiacetal covalent intermediate with the aldehyde substrate. The substrate specificity of the enzyme is broad and is usually dependent on its subcellular localization.In mammalian systems, three classes of aldehyde dehydrogenases have been identified on the basis of amino acid sequence and subReprint requests to:

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A Structural Explanation for the Retinal Specificity of Class 1 ALDH Enzymes

Moore¹,

Baker

Blythe³

et al. 1999

Advances in Experimental Medicine and Biology

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Crystallization and Preliminary X-ray Diffraction Studies on Cytosolic (Class 1) Aldehyde Dehydrogenase from Sheep Liver

Cited by 6 publications

References 0 publications

Sheep liver cytosolic aldehyde dehydrogenase: the structure reveals the basis for the retinal specificity of class 1 aldehyde dehydrogenases

Sheep liver cytosolic aldehyde dehydrogenase: the structure reveals the basis for the retinal specificity of class 1 aldehyde dehydrogenases

Crystallization and preliminary X‐ray analysis of aldehyde dehydrogenase from Vibrio harveyi

A Structural Explanation for the Retinal Specificity of Class 1 ALDH Enzymes

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