PMID- 10631996
OWN - NLM
STAT- MEDLINE
DCOM- 20000214
LR  - 20181113
IS  - 0961-8368 (Print)
IS  - 0961-8368 (Linking)
VI  - 8
IP  - 12
DP  - 1999 Dec
TI  - Human liver mitochondrial aldehyde dehydrogenase: three-dimensional structure and
      the restoration of solubility and activity of chimeric forms.
PG  - 2784-90
AB  - Human liver cytosolic and mitochondrial isozymes of aldehyde dehydrogenase share 
      70% sequence identity. However, the first 21 residues are not conserved between
      the human isozymes (15% identity). The three-dimensional structures of the beef
      mitochondrial and sheep cytosolic forms have virtually identical
      three-dimensional structures. Here, we solved the structure of the human
      mitochondrial enzyme and found it to be identical to the beef enzyme. The first
      21 residues are found on the surface of the enzyme and make no contact with other
      subunits in the tetramer. A pair of chimeric enzymes between the human isozymes
      was made. Each chimera had the first 21 residues from one isozyme and the
      remaining 479 from the other. When the first 21 residues were from the
      mitochondrial isozyme, an enzyme with cytosolic-like properties was produced. The
      other was expressed but was insoluble. It was possible to restore solubility and 
      activity to the chimera that had the first 21 cytosolic residues fused to the
      mitochondrial ones by making point mutations to residues at the N-terminal end.
      When residue 19 was changed from tyrosine to a cysteine, the residue found in the
      mitochondrial form, an active enzyme could be made though the Km for NAD+ was 35 
      times higher than the native mitochondrial isozyme and the specific activity was 
      reduced by 75%. This residue interacts with residue 203, a nonconserved,
      nonactive site residue. A mutation of residue 18, which also interacts with 203, 
      restored solubility, but not activity. Mutation to residue 15, which interacts
      with 104, also restored solubility but not activity. It appears that to have a
      soluble or active enzyme a favorable interaction must occur between a residue in 
      a surface loop and a residue elsewhere in the molecule even though neither make
      contact with the active site region of the enzyme.
FAU - Ni, L
AU  - Ni L
AD  - Department of Biochemistry, Purdue University, West Lafayette, Indiana
      47907-1153, USA.
FAU - Zhou, J
AU  - Zhou J
FAU - Hurley, T D
AU  - Hurley TD
FAU - Weiner, H
AU  - Weiner H
LA  - eng
SI  - PDB/1CW3
GR  - AA05812/AA/NIAAA NIH HHS/United States
GR  - AA11982/AA/NIAAA NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Protein Sci
JT  - Protein science : a publication of the Protein Society
JID - 9211750
RN  - 0 (Isoenzymes)
RN  - EC 1.2.1.3 (Aldehyde Dehydrogenase)
SB  - IM
MH  - Aldehyde Dehydrogenase/*chemistry
MH  - Crystallography, X-Ray
MH  - Fluorescence
MH  - Humans
MH  - Isoenzymes/chemistry
MH  - Mitochondria, Liver/*chemistry
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Solubility
PMC - PMC2144226
EDAT- 2000/01/13 09:00
MHDA- 2000/02/19 09:00
CRDT- 2000/01/13 09:00
PHST- 2000/01/13 09:00 [pubmed]
PHST- 2000/02/19 09:00 [medline]
PHST- 2000/01/13 09:00 [entrez]
AID - 10.1110/ps.8.12.2784 [doi]
PST - ppublish
SO  - Protein Sci. 1999 Dec;8(12):2784-90. doi: 10.1110/ps.8.12.2784.