PMID- 10231530
OWN - NLM
STAT- MEDLINE
DCOM- 19990601
LR  - 20131121
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 38
IP  - 18
DP  - 1999 May 4
TI  - Biochemical characterization and crystal structure determination of human heart
      short chain L-3-hydroxyacyl-CoA dehydrogenase provide insights into catalytic
      mechanism.
PG  - 5786-98
AB  - Human heart short chain L-3-hydroxyacyl-CoA dehydrogenase (SCHAD) catalyzes the
      oxidation of the hydroxyl group of L-3-hydroxyacyl-CoA to a keto group,
      concomitant with the reduction of NAD+ to NADH, as part of the beta-oxidation
      pathway. The homodimeric enzyme has been overexpressed in Escherichia coli,
      purified to homogeneity, and studied using biochemical and crystallographic
      techniques. The dissociation constants of NAD+ and NADH have been determined over
      a broad pH range and indicate that SCHAD binds reduced cofactor preferentially.
      Examination of apparent catalytic constants reveals that SCHAD displays optimal
      enzymatic activity near neutral pH, with catalytic efficiency diminishing rapidly
      toward pH extremes. The crystal structure of SCHAD complexed with NAD+ has been
      solved using multiwavelength anomalous diffraction techniques and a
      selenomethionine-substituted analogue of the enzyme. The subunit structure is
      comprised of two domains. The first domain is similar to other alpha/beta
      dinucleotide folds but includes an unusual helix-turn-helix motif which extends
      from the central beta-sheet. The second, or C-terminal, domain is primarily
      alpha-helical and mediates subunit dimerization and, presumably,
      L-3-hydroxyacyl-CoA binding. Molecular modeling studies in which
      L-3-hydroxybutyryl-CoA was docked into the enzyme-NAD+ complex suggest that His
      158 serves as a general base, abstracting a proton from the 3-OH group of the
      substrate. Furthermore, the ability of His 158 to perform such a function may be 
      enhanced by an electrostatic interaction with Glu 170, consistent with previous
      biochemical observations. These studies provide further understanding of the
      molecular basis of several inherited metabolic disease states correlated with
      L-3-hydroxyacyl-CoA dehydrogenase deficiencies.
FAU - Barycki, J J
AU  - Barycki JJ
AD  - Department of Biochemistry, Molecular Biology, and Biophysics, University of
      Minnesota, Minneapolis 55455, USA.
FAU - O'Brien, L K
AU  - O'Brien LK
FAU - Bratt, J M
AU  - Bratt JM
FAU - Zhang, R
AU  - Zhang R
FAU - Sanishvili, R
AU  - Sanishvili R
FAU - Strauss, A W
AU  - Strauss AW
FAU - Banaszak, L J
AU  - Banaszak LJ
LA  - eng
SI  - PDB/2HDH
SI  - PDB/3HAD
GR  - 1F32-DK09759-01/DK/NIDDK NIH HHS/United States
GR  - GM13925/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Recombinant Proteins)
RN  - 0U46U6E8UK (NAD)
RN  - EC 1.1.1.- (3-Hydroxyacyl CoA Dehydrogenases)
SB  - IM
MH  - 3-Hydroxyacyl CoA Dehydrogenases/*chemistry/genetics/isolation & purification
MH  - Amino Acid Sequence
MH  - Binding Sites
MH  - Catalysis
MH  - Crystallization
MH  - Crystallography, X-Ray
MH  - Dimerization
MH  - Escherichia coli/genetics
MH  - Humans
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Myocardium/*enzymology
MH  - NAD/chemistry
MH  - Protein Structure, Tertiary
MH  - Recombinant Proteins/biosynthesis/chemistry/isolation & purification
EDAT- 1999/05/08 00:00
MHDA- 1999/05/08 00:01
CRDT- 1999/05/08 00:00
PHST- 1999/05/08 00:00 [pubmed]
PHST- 1999/05/08 00:01 [medline]
PHST- 1999/05/08 00:00 [entrez]
AID - 10.1021/bi9829027 [doi]
AID - bi9829027 [pii]
PST - ppublish
SO  - Biochemistry. 1999 May 4;38(18):5786-98. doi: 10.1021/bi9829027.