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Lee McAlister-Henn, Professor;
Ph.D., UT Southwestern Medical Center, 1980.
Telephone: (210) 567-3782 E-mail:
henn@uthscsa.edu
Allosteric Regulation of Energy
Metabolism and Cellular Sources of NADPH for Antioxidant Function
Research in this laboratory
uses molecular genetic approaches to analyze regulation of the intracellular
redox environment. (A) A major area of investigation is the regulation
of rates of oxidative metabolism at the level of allosteric regulation
of mitochondrial NAD+-specific isocitrate dehydrogenase (IDH), which catalyzes
a rate-limiting step in the TCA cycle. We are constructing mutant
forms of the enzyme in yeast to investigate the extent and importance of
allosteric regulation in vivo. We are also investigating the X-ray
crystallographic structure of the enzyme (see Figure), which serves as
an excellent model for the co-evolution of regulatory and catalytic ligand-binding
sites and for fundamental principles involved in allostery. [This
work is supported by NIH grant GM051265.]
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IDH is an octameric enzyme composed of four regulatory
subunits (gold) and four catalytic subunits (navy). |
(B) In another major area, we
focus on two essential cytosolic sources of NADPH (the hexose monophosphate
pathway and cytosolic NADP+-specific isocitrate dehydrogenase). Loss
of both these sources results in a rapid decrease in viability of yeast
cells under conditions when metabolic flux through the pathways of peroxisomal
?-oxidation or of mitochondrial respiration is increased. This decrease
in viability is due to the accumulation of deleterious byproducts of these
endogenous metabolic pathways due to inadequate production of NADPH for
protective antioxidant enzyme systems. We are determining the endogenous
macromolecular targets of endogenous oxidative metabolic byproducts and
their effects on the process of aging in yeast, and we are extending these
observations to mammalian systems. [This work is supported by NIH
grant AG017477.] (C) In both these areas of investigation, we have
developed techniques to quantify levels of various cellular metabolites
and of reduced and oxidized forms of NAD(P)+ cofactors. These measurements
have led to an understanding of dramatic and rapid changes that can occur
in the cellular redox environment, and they have led to major new hypotheses
related to signaling events that control rates of flux through central
metabolic pathways.
Trainees participate in
all aspects of these research projects. Each trainee is encouraged
to choose hypotheses of interest and is expected to develop the experimental
expertise with a variety of methods (from classical to molecular genetics,
mutagenesis, protein expression and purification, kinetic/structural analyses,
physiological assays, etc.) as needed to address his/her goals
Hu
G, Taylor AB, McAlister-Henn L, Hart PJ.
Crystal structure of the yeast nicotinamidase Pnc1p.
Arch Biochem Biophys. 2007 May 1;461(1):66-75. Epub 2007 Mar
2.
PMID: 17382284
Hu
G, McAlister-Henn L.
Novel allosteric properties produced by residue substitutions in the
subunit interface of yeast NAD+-specific isocitrate dehydrogenase.
Arch Biochem Biophys. 2006 Sep 15;453(2):207-16. Epub 2006 Jul
12.
PMID: 16884682
Hu
G, Lin AP, McAlister-Henn L.
Physiological consequences of loss of allosteric activation of yeast
NAD+-specific isocitrate dehydrogenase.
J Biol Chem. 2006 Jun 23;281(25):16935-42. Epub 2006 Apr 18.
PMID: 16621803
Anderson
SL, Lin AP, McAlister-Henn L.
Analysis of interactions with mitochondrial mRNA using mutant forms
of yeast NAD(+)-specific isocitrate dehydrogenase.
Biochemistry. 2005 Dec 20;44(50):16776-84.
PMID: 16342968
Hu
G, Taylor AB, McAlister-Henn L, Hart PJ.
Crystallization and preliminary X-ray crystallographic analysis of
yeast NAD+-specific isocitrate dehydrogenase.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 May 1;61(Pt
5):486-8. Epub 2005 Apr 22. Erratum in: Acta Crystallograph Sect F Struct
Biol Cryst Commun. 2006 Jul 1;62(Pt 7):709.
PMID: 16511075
Minard
KI, McAlister-Henn L.
Sources of NADPH in yeast vary with carbon source.
J Biol Chem. 2005 Dec 2;280(48):39890-6. Epub 2005 Sep 22.
PMID: 16179340
Contreras-Shannon
V, Lin AP, McCammon MT, McAlister-Henn L.
Kinetic properties and metabolic contributions of yeast mitochondrial
and cytosolic NADP+-specific isocitrate dehydrogenases.
J Biol Chem. 2005 Feb 11;280(6):4469-75. Epub 2004 Dec 1.
PMID: 15574419 [PubMed - indexed for MEDLINE]
Contreras-Shannon
V, McAlister-Henn L.
Influence of compartmental localization on the function of yeast NADP+-specific
isocitrate dehydrogenases.
Arch Biochem Biophys. 2004 Mar 15;423(2):235-46.
PMID: 15001388 [PubMed - indexed for MEDLINE]
McCammon
MT, McAlister-Henn L.
Multiple cellular consequences of isocitrate dehydrogenase isozyme
dysfunction.
Arch Biochem Biophys. 2003 Nov 15;419(2):222-33.
PMID: 14592466 [PubMed - indexed for MEDLINE]
Anoop
VM, Basu U, McCammon MT, McAlister-Henn L, Taylor GJ.
Modulation of citrate metabolism alters aluminum tolerance in yeast
and transgenic canola overexpressing a mitochondrial citrate synthase.
Plant Physiol. 2003 Aug;132(4):2205-17.
PMID: 12913175 [PubMed - indexed for MEDLINE]
Gibson
N, McAlister-Henn L.
Physical and genetic interactions of cytosolic malate dehydrogenase
with other gluconeogenic enzymes.
J Biol Chem. 2003 Jul 11;278(28):25628-36. Epub 2003 May 01.
PMID: 12730240 [PubMed - indexed for MEDLINE]
McCammon
MT, Epstein CB, Przybyla-Zawislak B, McAlister-Henn L, Butow RA.
Global transcription analysis of Krebs tricarboxylic acid cycle mutants
reveals an alternating pattern of gene expression and effects on hypoxic
and oxidative genes.
Mol Biol Cell. 2003 Mar;14(3):958-72.
PMID: 12631716 [PubMed - indexed for MEDLINE]
Lin
AP, McAlister-Henn L.
Homologous binding sites in yeast isocitrate dehydrogenase for cofactor
(NAD+) and allosteric activator (AMP).
J Biol Chem. 2003 Apr 11;278(15):12864-72. Epub 2003 Jan 31.
PMID: 12562755 [PubMed - indexed for MEDLINE]
Anderson
SL, Schirf V, McAlister-Henn L.
Effect of AMP on mRNA binding by yeast NAD+-specific isocitrate dehydrogenase.
Biochemistry. 2002 Jun 4;41(22):7065-73.
PMID: 12033940 [PubMed - indexed for MEDLINE]
Lin
AP, McAlister-Henn L.
Isocitrate binding at two functionally distinct sites in yeast NAD+-specific
isocitrate dehydrogenase.
J Biol Chem. 2002 Jun 21;277(25):22475-83. Epub 2002 Apr 12.
PMID: 11953438 [PubMed - indexed for MEDLINE]
Lin
AP, McCammon MT, McAlister-Henn L.
Kinetic and physiological effects of alterations in homologous isocitrate-binding
sites of yeast NAD(+)-specific isocitrate dehydrogenase.
Biochemistry. 2001 Nov 27;40(47):14291-301.
PMID: 11714283 [PubMed - indexed for MEDLINE]
Minard
KI, McAlister-Henn L.
Antioxidant function of cytosolic sources of NADPH in yeast.
Free Radic Biol Med. 2001 Sep 15;31(6):832-43.
PMID: 11557322 [PubMed - indexed for MEDLINE]
Panisko
EA, McAlister-Henn L.
Subunit interactions of yeast NAD+-specific isocitrate dehydrogenase.
J Biol Chem. 2001 Jan 12;276(2):1204-10.
PMID: 11042198 [PubMed - indexed for MEDLINE]
Anderson
SL, Minard KI, McAlister-Henn L.
Allosteric inhibition of NAD+-specific isocitrate dehydrogenase by
a mitochondrial mRNA.
Biochemistry. 2000 May 16;39(19):5623-9.
PMID: 10801312 [PubMed - indexed for MEDLINE]
McAlister-Henn
L, Gibson N, Panisko E.
Applications of the yeast two-hybrid system.
Methods. 1999 Oct;19(2):330-7.
PMID: 10527735 [PubMed - indexed for MEDLINE]
Minard
KI, McAlister-Henn L.
Dependence of peroxisomal beta-oxidation on cytosolic sources of NADPH.
J Biol Chem. 1999 Feb 5;274(6):3402-6.
PMID: 9920883 [PubMed - indexed for MEDLINE]
Minard
KI, Jennings GT, Loftus TM, Xuan D, McAlister-Henn L.
Sources of NADPH and expression of mammalian NADP+-specific isocitrate
dehydrogenases in Saccharomyces cerevisiae.
J Biol Chem. 1998 Nov 20;273(47):31486-93.
PMID: 9813062 [PubMed - indexed for MEDLINE]
Small
WC, McAlister-Henn L.
Identification of a cytosolically directed NADH dehydrogenase in mitochondria
of Saccharomyces cerevisiae.
J Bacteriol. 1998 Aug;180(16):4051-5.
PMID: 9696750 [PubMed - indexed for MEDLINE]
Jennings
GT, Minard KI, McAlister-Henn L.
Expression and mutagenesis of mammalian cytosolic NADP+-specific isocitrate
dehydrogenase.
Biochemistry. 1997 Nov 4;36(44):13743-7.
PMID: 9354646 [PubMed - indexed for MEDLINE]
Zhao
WN, McAlister-Henn L.
Affinity purification and kinetic analysis of mutant forms of yeast
NAD+-specific isocitrate dehydrogenase.
J Biol Chem. 1997 Aug 29;272(35):21811-7.
PMID: 9268311 [PubMed - indexed for MEDLINE]
Small
WC, McAlister-Henn L.
Metabolic effects of altering redundant targeting signals for yeast
mitochondrial malate dehydrogenase.
Arch Biochem Biophys. 1997 Aug 1;344(1):53-60.
PMID: 9244381 [PubMed - indexed for MEDLINE]
McAlister-Henn
L, Small WC.
Molecular genetics of yeast TCA cycle isozymes.
Prog Nucleic Acid Res Mol Biol. 1997;57:317-39. Review. No abstract
available.
PMID: 9175438 [PubMed - indexed for MEDLINE]
Soundar
S, Jennings GT, McAlister-Henn L, Colman RF.
Expression of pig heart mitochondrial NADP-dependent isocitrate dehydrogenase
in Escherichia coli.
Protein Expr Purif. 1996 Nov;8(3):305-12.
PMID: 8936592 [PubMed - indexed for MEDLINE]
Zhao
WN, McAlister-Henn L.
Expression and gene disruption analysis of the isocitrate dehydrogenase
family in yeast.
Biochemistry. 1996 Jun 18;35(24):7873-8.
PMID: 8672488 [PubMed - indexed for MEDLINE]
Zhao
WN, McAlister-Henn L.
Assembly and function of a cytosolic form of NADH-specific isocitrate
dehydrogenase in yeast.
J Biol Chem. 1996 Apr 26;271(17):10347-52.
PMID: 8626605 [PubMed - indexed for MEDLINE]
McAlister-Henn
L, Steffan JS, Minard KI, Anderson SL.
Expression and function of a mislocalized form of peroxisomal malate
dehydrogenase (MDH3) in yeast.
J Biol Chem. 1995 Sep 8;270(36):21220-5.
PMID: 7673155 [PubMed - indexed for MEDLINE]
Huang
YC, Haselbeck RJ, McAlister-Henn L, Colman RF.
N-ethylmaleimide profiling of yeast NADP-dependent isocitrate dehydrogenase.
Arch Biochem Biophys. 1995 Jan 10;316(1):485-92.
PMID: 7840654 [PubMed - indexed for MEDLINE]
Minard
KI, McAlister-Henn L.
Glucose-induced phosphorylation of the MDH2 isozyme of malate dehydrogenase
in Saccharomyces cerevisiae.
Arch Biochem Biophys. 1994 Dec;315(2):302-9.
PMID: 7986072 [PubMed - indexed for MEDLINE]
Jennings
GT, Sechi S, Stevenson PM, Tuckey RC, Parmelee D, McAlister-Henn L.
Cytosolic NADP(+)-dependent isocitrate dehydrogenase. Isolation of
rat cDNA and study of tissue-specific and developmental expression of mRNA.
J Biol Chem. 1994 Sep 16;269(37):23128-34.
PMID: 8083215 [PubMed - indexed for MEDLINE]
Loftus
TM, Hall LV, Anderson SL, McAlister-Henn L.
Isolation, characterization, and disruption of the yeast gene encoding
cytosolic NADP-specific isocitrate dehydrogenase.
Biochemistry. 1994 Aug 16;33(32):9661-7.
PMID: 8068643 [PubMed - indexed for MEDLINE]
Cupp
JR, McAlister-Henn L.
Kinetic analysis of NAD(+)-isocitrate dehydrogenase with altered isocitrate
binding sites: contribution of IDH1 and IDH2 subunits to regulation and
catalysis.
Biochemistry. 1993 Sep 14;32(36):9323-8.
PMID: 8369302 [PubMed - indexed for MEDLINE]
Haselbeck
RJ, McAlister-Henn L.
Function and expression of yeast mitochondrial NAD- and NADP-specific
isocitrate dehydrogenases.
J Biol Chem. 1993 Jun 5;268(16):12116-22.
PMID: 8099357 [PubMed - indexed for MEDLINE]
Steffan
JS, McAlister-Henn L.
Isolation and characterization of the yeast gene encoding the MDH3
isozyme of malate dehydrogenase.
J Biol Chem. 1992 Dec 5;267(34):24708-15.
PMID: 1447211 [PubMed - indexed for MEDLINE]
Sumegi
B, McCammon MT, Sherry AD, Keys DA, McAlister-Henn L, Srere PA.
Metabolism of [3-13C]pyruvate in TCA cycle mutants of yeast.
Biochemistry. 1992 Sep 22;31(37):8720-5.
PMID: 1390657 [PubMed - indexed for MEDLINE]
Minard
KI, McAlister-Henn L.
Glucose-induced degradation of the MDH2 isozyme of malate dehydrogenase
in yeast.
J Biol Chem. 1992 Aug 25;267(24):17458-64.
PMID: 1324938 [PubMed - indexed for MEDLINE]
Cupp
JR, McAlister-Henn L.
Cloning and characterization of the gene encoding the IDH1 subunit
of NAD(+)-dependent isocitrate dehydrogenase from Saccharomyces cerevisiae.
J Biol Chem. 1992 Aug 15;267(23):16417-23.
PMID: 1644826 [PubMed - indexed for MEDLINE]
Haselbeck
RJ, Colman RF, McAlister-Henn L.
Isolation and sequence of a cDNA encoding porcine mitochondrial NADP-specific
isocitrate dehydrogenase.
Biochemistry. 1992 Jul 14;31(27):6219-23.
PMID: 1627563 [PubMed - indexed for MEDLINE]
Said
HM, McAlister-Henn L, Mohammadkhani R, Horne DW.
Uptake of biotin by isolated rat liver mitochondria.
Am J Physiol. 1992 Jul;263(1 Pt 1):G81-6.
PMID: 1636720 [PubMed - indexed for MEDLINE]
Steffan
JS, Minard KI, McAlister-Henn L.
Expression and function of heterologous forms of malate dehydrogenase
in yeast.
Arch Biochem Biophys. 1992 Feb 14;293(1):93-102.
PMID: 1731644 [PubMed - indexed for MEDLINE]
Cupp
JR, McAlister-Henn L.
NAD(+)-dependent isocitrate dehydrogenase. Cloning, nucleotide sequence,
and disruption of the IDH2 gene from Saccharomyces cerevisiae.
J Biol Chem. 1991 Nov 25;266(33):22199-205.
PMID: 1939242 [PubMed - indexed for MEDLINE]
Steffan
JS, McAlister-Henn L.
Structural and functional effects of mutations altering the subunit
interface of mitochondrial malate dehydrogenase.
Arch Biochem Biophys. 1991 Jun;287(2):276-82.
PMID: 1898005 [PubMed - indexed for MEDLINE]
Haselbeck
RJ, McAlister-Henn L.
Isolation, nucleotide sequence, and disruption of the Saccharomyces
cerevisiae gene encoding mitochondrial NADP(H)-specific isocitrate dehydrogenase.
J Biol Chem. 1991 Feb 5;266(4):2339-45.
PMID: 1989987 [PubMed - indexed for MEDLINE]
Minard
KI, McAlister-Henn L.
Isolation, nucleotide sequence analysis, and disruption of the MDH2
gene from Saccharomyces cerevisiae: evidence for three isozymes of yeast
malate dehydrogenase.
Mol Cell Biol. 1991 Jan;11(1):370-80.
PMID: 1986231 [PubMed - indexed for MEDLINE]
Keys
DA, McAlister-Henn L.
Subunit structure, expression, and function of NAD(H)-specific isocitrate
dehydrogenase in Saccharomyces cerevisiae.
J Bacteriol. 1990 Aug;172(8):4280-7.
PMID: 2198251 [PubMed - indexed for MEDLINE]
Thompson
LM, McAlister-Henn L.
Dispensable presequence for cellular localization and function of mitochondrial
malate dehydrogenase from Saccharomyces cerevisiae.
J Biol Chem. 1989 Jul 15;264(20):12091-6.
PMID: 2663847 [PubMed - indexed for MEDLINE]
Thompson
LM, Sutherland P, Steffan JS, McAlister-Henn L.
Gene sequence and primary structure of mitochondrial malate dehydrogenase
from Saccharomyces cerevisiae.
Biochemistry. 1988 Nov 1;27(22):8393-400.
PMID: 3072021 [PubMed - indexed for MEDLINE]
McAlister-Henn
L.
Evolutionary relationships among the malate dehydrogenases.
Trends Biochem Sci. 1988 May;13(5):178-81. No abstract available.
PMID: 3076279 [PubMed - indexed for MEDLINE]
McAlister-Henn
L, Thompson LM.
Isolation and expression of the gene encoding yeast mitochondrial malate
dehydrogenase.
J Bacteriol. 1987 Nov;169(11):5157-66.
PMID: 3312168 [PubMed - indexed for MEDLINE]
McAlister-Henn
L, Blaber M, Bradshaw RA, Nisco SJ.
Complete nucleotide sequence of the Escherichia coli gene encoding
malate dehydrogenase.
Nucleic Acids Res. 1987 Jun 25;15(12):4993. No abstract available.
PMID: 3299262 [PubMed - indexed for MEDLINE]
Sutherland
P, McAlister-Henn L.
Isolation and expression of the Escherichia coli gene encoding malate
dehydrogenase.
J Bacteriol. 1985 Sep;163(3):1074-9.
PMID: 2993232 [PubMed - indexed for MEDLINE]
McAlister
L, Holland MJ.
Differential expression of the three yeast glyceraldehyde-3-phosphate
dehydrogenase genes.
J Biol Chem. 1985 Dec 5;260(28):15019-27.
PMID: 3905788 [PubMed - indexed for MEDLINE]
McAlister
L, Holland MJ.
Isolation and characterization of yeast strains carrying mutations
in the glyceraldehyde-3-phosphate dehydrogenase genes.
J Biol Chem. 1985 Dec 5;260(28):15013-8.
PMID: 2999100 [PubMed - indexed for MEDLINE]
Finkelstein
DB, Strausberg S, McAlister L.
Alterations of transcription during heat shock of Saccharomyces cerevisiae.
J Biol Chem. 1982 Jul 25;257(14):8405-11.
PMID: 6177697 [PubMed - indexed for MEDLINE]
McAlister
L, Holland MJ.
Targeted deletion of a yeast enolase structural gene. Identification
and isolation of yeast enolase isozymes.
J Biol Chem. 1982 Jun 25;257(12):7181-8.
PMID: 6282834 [PubMed - indexed for MEDLINE]
McAlister
LE, Evans EL, Smith TE.
Properties of a mutant Escherichia coli phosphoenolpyruvate carboxylase
deficient in coregulation by intermediary metabolites.
J Bacteriol. 1981 Apr;146(1):200-8.
PMID: 7012114 [PubMed - indexed for MEDLINE]
Finkelstein
DB, McAlister L.
alpha-Factor-mediatd modification of a 32P-labeled protein by MATa
cells of Saccharomyces cerevisiae.
J Biol Chem. 1981 Mar 10;256(5):2561-6.
PMID: 7007388 [PubMed - indexed for MEDLINE]
McAlister
L, Finkelstein DB.
Alterations in translatable ribonucleic acid after heat shock of Saccharomyces
cerevisiae.
J Bacteriol. 1980 Aug;143(2):603-12.
PMID: 7009554 [PubMed - indexed for MEDLINE]
McAlister
L, Finkelstein DB.
Heat shock proteins and thermal resistance in yeast.
Biochem Biophys Res Commun. 1980 Apr 14;93(3):819-24. No abstract
available.
PMID: 6992776 [PubMed - indexed for MEDLINE]
McAlister
LE, Allison VF, Jeter JR, Nations C.
High-density induction of a quiescent cell state in Physarum polycephalum.
J Cell Sci. 1977 Jun;25:179-90.
PMID: 561094 [PubMed - indexed for MEDLINE]
