PMID- 10625657
OWN - NLM
STAT- MEDLINE
DCOM- 20000218
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 275
IP  - 2
DP  - 2000 Jan 14
TI  - Expression, purification, and characterization of natural mutants of human
      aldolase B. Role of quaternary structure in catalysis.
PG  - 1145-51
AB  - Fructaldolases (EC 4.1.2.13) are ancient enzymes of glycolysis that catalyze the 
      reversible cleavage of phosphofructose esters into cognate triose (phosphates).
      Three vertebrate isozymes of Class I aldolase have arisen by gene duplication and
      display distinct activity profiles with fructose 1,6-bisphosphate and with
      fructose 1-phosphate. We describe the biochemical and biophysical
      characterization of seven natural human aldolase B variants, identified in
      patients suffering from hereditary fructose intolerance and expressed as
      recombinant proteins in E. coli, from which they were purified to homogeneity.
      The mutant aldolases were all missense variants and could be classified into two 
      principal groups: catalytic mutants, with retained tetrameric structure but
      altered kinetic properties (W147R, R303W, and A337V), and structural mutants, in 
      which the homotetramers readily dissociate into subunits with greatly impaired
      enzymatic activity (A149P, A174D, L256P, and N334K). Investigation of these two
      classes of mutant enzyme suggests that the integrity of the quaternary structure 
      of aldolase B is critical for maintaining its full catalytic function.
FAU - Rellos, P
AU  - Rellos P
AD  - Department of Medicine, University of Cambridge, Level 5, Addenbrooke's Hospital,
      Cambridge CB2 2QQ, United Kingdom.
FAU - Sygusch, J
AU  - Sygusch J
FAU - Cox, T M
AU  - Cox TM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Recombinant Proteins)
RN  - EC 4.1.2.13 (Fructose-Bisphosphate Aldolase)
SB  - IM
MH  - Amino Acid Substitution
MH  - Animals
MH  - Catalysis
MH  - Chromatography, Ion Exchange
MH  - Cloning, Molecular
MH  - Escherichia coli
MH  - Fructose-Bisphosphate Aldolase/*chemistry/*genetics/isolation &
      purification/metabolism
MH  - *Genetic Variation
MH  - Humans
MH  - Kinetics
MH  - Liver/enzymology
MH  - Models, Molecular
MH  - Mutagenesis, Site-Directed
MH  - *Mutation, Missense
MH  - Protein Structure, Quaternary
MH  - Rabbits
MH  - Recombinant Proteins/chemistry/isolation & purification/metabolism
EDAT- 2000/01/08 09:00
MHDA- 2000/02/26 09:00
CRDT- 2000/01/08 09:00
PHST- 2000/01/08 09:00 [pubmed]
PHST- 2000/02/26 09:00 [medline]
PHST- 2000/01/08 09:00 [entrez]
AID - 10.1074/jbc.275.2.1145 [doi]
PST - ppublish
SO  - J Biol Chem. 2000 Jan 14;275(2):1145-51. doi: 10.1074/jbc.275.2.1145.