PDBsum entry 1mgo

Oxidoreductase

PDB id: 1mgo

Contents

Protein chains

374 a.a. *

Ligands

NAD ×2

PFB ×2

MPD

Metals

_ZN ×4

Waters ×465

* Residue conservation analysis

References listed in PDB file

Key reference

Title

Mobility of fluorobenzyl alcohols bound to liver alcohol dehydrogenases as determined by nmr and X-Ray crystallographic studies.

Authors

J.K.Rubach, B.V.Plapp.

Ref.

Biochemistry, 2002, 41, 15770-15779. [DOI no: 10.1021/bi026581h]

PubMed id

12501206

Abstract

The relationship between substrate mobility and catalysis was studied with wild-type and Phe93Ala (F93A) horse liver alcohol dehydrogenase (ADH). Wild-type ADH binds 2,3,4,5,6-pentafluorobenzyl alcohol in one position as shown by X-ray results, and (19)F NMR shows five resonances for the fluorines of the bound alcohol. The two meta-fluorines exchange positions with a rate constant of about 4 s(-1), indicating that mobility (ring flipping) of the benzyl alcohol is relatively restricted. The wild-type enzyme binds 2,3-difluorobenzyl alcohol in two alternative conformations that are related by a ring flip and a small translation of the fluorinated benzene ring, and the (19)F NMR spectrum shows three resonances for the two bound fluorines, consistent with the two orientations. Phe-93 interacts with the bound benzyl alcohols, and the F93A substitution decreases the rate constants for hydride transfer for benzyl alcohol oxidation and benzaldehyde reduction by 7.4- and 130-fold, respectively. The structure of F93A ADH crystallized with NAD(+) and 2,3,4,5,6-pentafluorobenzyl alcohol is similar to the structure of the wild-type enzyme complex except that the pentafluorobenzyl alcohol is not found in one position. The (19)F NMR spectrum of the F93A ADH-NAD(+)-pentafluorobenzyl alcohol complex shows three resonances for the bound fluorines. Line shape analysis of the spectrum suggests the bound pentafluorobenzyl ring undergoes rapid ring-flipping at about 20 000 s(-1). The F93A substitution greatly increases the mobility of the benzyl alcohol but modestly and differentially decreases the probability that the substrate is preorganized for hydride transfer.

Secondary reference #1

Title

Structures of horse liver alcohol dehydrogenase complexed with NAD+ and substituted benzyl alcohols.

Authors

S.Ramaswamy, H.Eklund, B.V.Plapp.

Ref.

Biochemistry, 1994, 33, 5230-5237. [DOI no: 10.1021/bi00183a028]

PubMed id

8172897

Secondary reference #2

Title

Binding of substrate in a ternary complex of horse liver alcohol dehydrogenase.

Authors

H.Eklund, B.V.Plapp, J.P.Samama, C.I.Brändén.

Ref.

J Biol Chem, 1982, 257, 14349-14358.

PubMed id

6754727

Secondary reference #3

Title

Structure of a triclinic ternary complex of horse liver alcohol dehydrogenase at 2.9 a resolution.

Authors

H.Eklund, J.P.Samma, L.Wallén, C.I.Brändén, A.Akeson, T.A.Jones.

Ref.

J Mol Biol, 1981, 146, 561-587. [DOI no: 10.1016/0022-2836(81)90047-4]

PubMed id

7024556

Figure 8.
PIG. 8. Diagrams illustrating interactins between NADH and LADHaue (a) The adenine ring in its electron density sandwiched between the side-chains of Ile224 and Ile269 in parts of their eletron densites. (b) Interactions between protein side-rhains and the ADP-ribose part of NAD. (v) Surrounding of th nicotinamide part of NAD. (d) Stereo diagram of Cn atoms of one subunit and bound coenzyme (in red).

Figure 10.
FIG. 10. Picture from the display system illustrating the interaction between the pyrophosphate moiety of NADH and helix aB of the roenzyme binding domain of LADHase.

The above figures are reproduced from the cited reference with permission from Elsevier

Secondary reference #4

Title

Three-Dimensional structure of horse liver alcohol dehydrogenase at 2-4 a resolution.

Authors

H.Eklund, B.Nordström, E.Zeppezauer, G.Söderlund, I.Ohlsson, T.Boiwe, B.O.Söderberg, O.Tapia, C.I.Brändén, A.Akeson.

Ref.

J Mol Biol, 1976, 102, 27-59.

PubMed id

178875