Michael J. McLeish
Australian National Research Fellowship, 1985-88; CSIRO Postdoctoral Fellowship, 1984-1985; La Trobe University Research Scholarship, 1979-1982.
Research
My primary research interests lie in the broadly defined area of mechanistic
enzymology. Two representative projects, which focus on distinctly different aspects
of this field, are as follows:
Inhibition of the epinephrine synthesizing enzyme, phenylethanolamine Nmethyltransferase
(PNMT).
While the role of peripheral epinephrine (adrenaline) is reasonably well understood its
role in the CNS is not so well defined. PNMT inhibitors could be used clarify its role,
particularly in the central control of hypertension. Unfortunately it is hard to get a
clear-cut picture of CNS epinephrine effects as most of the currently available
inhibitors also interact with adrenoceptors. We are using a combination of QSAR
methods, kinetic and mutagenic analysis as well as structure-based design to develop
potent and specific PNMT inhibitors. Initially these will be used to delineate the role of
CNS epinephrine but, ultimately, we hope they will be useful as antihypertensives
with a novel mode of action.
Structure and function of the mandelate pathway enzymes.
This pathway, which permits a variety of pseudomonads to use the enantiomers of
mandelic acid as sole carbon source, is thought to be a paradigm for the study of
functional evolution. The known pathway comprises five enzymes, each of which
utilizes quite different chemistry. We are using a combination X-ray crystallography,
site-direct mutagenesis and mechanistic studies to understand the chemistry behind
each of these enzymes. Armed with this knowledge we aim to use directed evolution
to develop a new pathway, the lactate pathway. In the new pathway, instead of
utilizing aromatic substrates, each enzyme will use the corresponding aliphatic
substrate. In some instances the evolutionary process should be relatively simple,
needing only minor changes in substrate specificity. In other cases the new enzyme
may need to employ new chemistry.
Recent Publications
G. Brandt, N. Nemeria, S. Chakraborty, M. J. McLeish, A. Yep, G. L. Kenyon, G.
Petsko, F. Jordan and D. Ringe "Probing the active center of benzaldehyde lyase with
substitutions and the pseudo-substrate analogue benzoylphosphonic acid methyl
ester" Biochemistry 2008, 47, 7734-7743.
S. Chakraborty, N. Nemeria, A. Yep, M. J. McLeish, G. L. Kenyon and F. Jordan "The
mechanism of benzaldehyde lyase studied via thiamin diphosphate-bound
Intermediates and kinetic isotope effects" Biochemistry 2008, 47, 3800-3809.
A. Yep, G. L. Kenyon and M. J. McLeish "Saturation mutagenesis of putative catalytic
residues of benzoylformate decarboxylase provides a challenge to the accepted
mechanism" Proceedings of the National Academy of Sciences USA 2008, 105, 5733-
5738.
C. K. Yeung, G. L. Kenyon and M. J. McLeish "Physical, kinetic and spectrophotometric
studies of a NAD(P)+-dependent benzaldehyde dehydrogenase from Pseudomonas
putida ATCC 12633" Biochimica et Biophysica Acta - Proteins and Proteomics 2008,
1248-1255.
N. Nemeria, L. Korotchkina, M. J. McLeish, G. L. Kenyon, M. S. Patel and F. Jordan
"Elucidation of the chemistry of enzyme-bound thiamin diphosphate prior to substrate
binding: defining internal equilibria among tautomeric and ionization states "
Biochemistry 2007, 46, 10739-10744.
C. Saehuan, T. Rojanarata, S. Wiyakrutta, M. J. McLeish and V. Meevootisom
"Isolation and characterization of a benzoylformate decarboxylase and a
NAD+/NADP+-dependent benzaldehyde dehydrogenase involved in D-phenylglycine
metabolism in Pseudomonas stutzeri ST-201" Biochimica et Biophysica Acta 2007,
1770, 1585-1592.
C. L. Gee, N. Drinkwater, J. D. A. Tyndall, G. L. Grunewald, Q. Qian Wu, M. J.
McLeish and J. L. Martin "Enzyme adaptation to inhibitor binding: A cryptic binding
site in phenylethanolamine N-methyltransferase" Journal of Medicinal Chemistry
2007, 50, 4845-4853.
A. K. Bera, L. S. Polovnikova, J. Roestamadji, T. S. Widlanski, G. L. Kenyon, M. J.
McLeish and M. S. Hasson "Mechanism-based inactivation of benzoylformate
decarboxylase, a thiamin diphosphate-dependent enzyme" Journal of the American
Chemical Society 2007, 129, 4120-4121.
E. Janzen, M. Müller, D. Kolter-Jung, M. M. Kneen, M. J. McLeish and M. Pohl
"Characterization of benzaldehyde lyase from Pseudomonas fluorescens - a versatile
enzyme for asymmetric C-C-bond formation" Bioorganic Chemistry 2006, 34, 345-
361.
M. J. McLeish and G. L. Kenyon "Using site-directed mutagenesis to elucidate
structure and mechanism in creatine kinase" In Creatine Kinase; Vial, C., Ed.; Nova
Science, 2006, pp. 27-68.
A. Yep, G. L. Kenyon and M. J. McLeish "Determinants of substrate specificity in
KdcA, a thiamin diphosphate-dependent decarboxylase" Bioorganic Chemistry 2006,
34, 325-336.
P.-F. Wang, A. J. Flynn, M. M. Naor, J. H. Jensen, G. Cui, K. M. Merz Jr., G. L.
Kenyon and M. J. McLeish "Exploring the role of the active site cysteine in human
muscle creatine kinase" Biochemistry 2006, 45, 11464-11472.
P.-F. Wang, G. L. Kenyon and M. J. McLeish "Heterogeneity of Escherichia coliexpressed
human muscle creatine kinase" IUBMB Life 2006, 58, 421-428.
A. D. Andricopulo, M. B. Akoachere, R. Krogh, C. Nickel, M. J. McLeish, G. L. Kenyon,
L. D. Arscott, C. H. J. Williams, E. Davioud-Charvet and K. Becker "Specific inhibitors
of Plasmodium falciparum thioredoxin reductase as potential antimalarial agents"
Bioorganic and Medicinal Chemistry Letters 2006, 16, 2283-2292.
C. L. Gee, J. D. A. Tyndall, G. L. Grunewald, Q. Wu, M. J. McLeish and J. L. Martin
"Binding mode of methyl acceptor substrates to the adrenaline-synthesizing enzyme
phenylethanolamine N-methyltransferase: Implications for catalysis" Biochemistry
2005, 44, 16875-16885.
M. M. Kneen, I. D. Pogozheva, G. L. Kenyon and M. J. McLeish "Exploring the active
site of benzaldehyde lyase by modeling and mutagenesis" Biochimica et Biophysica
Acta - Proteins and Proteomics 2005, 1753, 263-271.
Q. Wu, C. L. Gee, F. Lin, J. D. Tyndall, J. L. Martin, G. L. Grunewald and M. J.
McLeish "Structural, mutagenic and kinetic analysis of the binding of substrates and
inhibitors of human phenylethanolamine N-methyltransferase" Journal of Medicinal
Chemistry 2005, 48, 7243-7252.
P. Siegert, M. J. McLeish, M. Baumann, M. M. Kneen, G. L. Kenyon and M. Pohl
"Exchanging the substrate specificities of pyruvate decarboxylase from Zymomonas
mobilis and benzoylformate decarboxylase from Pseudomonas putida" Protein
Engineering, Design and Selection 2005, 18, 345-357.
P.-F. Wang, A. D. Flynn, M. J. McLeish and G. L. Kenyon "Loop movement and
catalysis in creatine kinase" IUBMB Life 2005, 57, 355-362.
M. J. McLeish and G. L. Kenyon "Relating structure to mechanism in creatine kinase"
Critical Reviews in Biochemistry and Molecular Biology 2005, 40, 1-20.v
C. L. Gee, A. Nourse, A.-Y. Hsin, Q. Wu, J. D. Tyndall, G. L. Grunewald, M. J. McLeish
and J. L. Martin "Disulfide-linked dimers of human adrenaline synthesizing enzyme
PNMT are catalytically active" Biochimica et Biophysica Acta - Proteins and Proteomics
2005, 1750, 82-92.
P. Siegert, M. Pohl, M. M. Kneen, I. D. Pogozheva, G. L. Kenyon and M. J. McLeish
"Exploring the substrate specificity of benzoylformate decarboxylase, pyruvate
decarboxylase and benzaldehyde lyase" In: Thiamin: Catalytic mechanisms and role in
normal and disease states; F. Jordan and M. S. Patel, Eds., Marcel Dekker Inc., 2004,
pp. 275-290.
Q. Wu, K. R. Criscione, L. Grunewald Gary and M. J. McLeish "Phenylethanolamine Nmethyltransferase:
re-evaluation of kinetic data" Bioorganic and Medicinal Chemistry
Letters 2004, 14, 4217-4220.
F. A. Romero, S. M. Vodonick, K. R. Criscione, M. J. McLeish and G. L. Grunewald
"Inhibitors of phenylethanolamine N-methyltransferase that are predicted to penetrate
the blood-brain barrier: Design, synthesis and evaluation of 3-fluoromethyl-7-(Nsubstituted-
aminosulfonyl)-1,2,3,4-tetrahydroisoquinolines that possess low affinity
towards the a2-adrenoceptor" Journal of Medicinal Chemistry 2004, 47, 4483-4493.
W. R. P. Novak, P.-F. Wang, M. J. McLeish, G. L. Kenyon and P. C. Babbitt "Isoleucine
69 and Valine 325 form a specificity pocket in human muscle creatine kinase"
Biochemistry 2004, 43, 13766-13774.
F. M. McMillan, J. Archbold, M. J. McLeish, J. M. Caine, K. R. Criscione, G. L.
Grunewald and J. L. Martin "Molecular recognition of sub-micromolar inhibitors by the
epinephrine-synthesising enzyme phenylethanolamine N-methyltransferase" Journal of
Medicinal Chemistry 2004, 47, 37-44.
K. N. Gopalakrishna, B. H. Stewart, M. M. Kneen, A. D. Andricopulo, G. L. Kenyon
and M. J. McLeish "Mandelamide hydrolase from Pseudomonas putida:
Characterization of a new member of the amidase signature family" Biochemistry
2004, 43, 7725-7735.
M. J. McLeish, G. L. Kenyon, E. S. Polovnikova, A. S. Bera, N. L. Anderson and M. S.
Hasson "Benzoylformate decarboxylase: Lessons in enzymology" In: Thiamin:
Catalytic mechanisms and role in normal and disease states; F. Jordan and M. S.
Patel, Eds., Marcel Dekker Inc., 2004, pp. 131-141.
E. S. Polovnikova, M. J. McLeish, E. A. Sergienko, J. T. Burgner, N. L. Anderson, A. K.
Bera, F. Jordan, G. L. Kenyon and M. S. Hasson "Structural and kinetic analysis of
catalysis by a thiamin diphosphate-dependent enzyme, benzoylformate
decarboxylase" Biochemistry 2003, 42, 1820-1830.
M. J. McLeish, M. M. Kneen, K. N. Gopalakrishna, C. W. Koo, P. C. Babbitt, J. A. Gerlt
and G. L. Kenyon "Identification and characterization of a mandelamide hydrolase and
an NAD(P)+-dependent benzaldehyde dehydrogenase from Pseudomonas putida ATCC
12633" Journal of Bacteriology 2003, 185, 2451-2456.
E. Davioud-Charvet, M. J. McLeish, D. M. Veine, D. Giegel, L. D. Arscott, A. D.
Andricopulo, K. Becker, S. Müller, R. H. Schirmer, C. H. Williams Jr. and G. L. Kenyon
"Mechanism-based inactivation of thioredoxin reductase by Mannich bases.
Implication for cytotoxicity" Biochemistry 2003, 42, 13319-13330.
M. J. McLeish and G. L. Kenyon "Approaches to the rational design of enzyme
inhibitors" In: Burger's Medicinal Chemistry 6th Ed., Vol. 1, Drug Discovery, Wiley and
Sons, 2003, pp. 715-779.
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