9 / 21
Page 51
conferenceseries
.com
Volume 8
Journal of Biotechnology & Biomaterials
ISSN: 2155-952X
Biotech Congress 2018 & Enzymology 2018
March 05-07, 2018
JOINT EVENT
20
th
Global Congress on
Biotechnology
3
rd
International Conference on
Enzymology and Molecular Biology
&
March 05-07, 2018 London, UK
N-acyltransferases and their role in fatty acid amide biosynthesis
David J Merkler
University of South Florida, USA
F
atty acid amides are a family of cell signaling lipids with the general structure of R-CO-NH-Y. This structural simplicity belies
a wealth of diversity amongst this lipid family as the R-group is derived from fatty acids (R-COOH) and the Y-group is derived
from biogenic amines (H2N-Y). The fatty acid amide family is divided into classes, defined by parent amines. Examples include the
N-acylethanolamines (NAEs, R-CO-NH-CH2-CH2OH) and the N-acylglycines (NAGs, R-CO-NH-CH2-COOH). Other classes of
fatty acid amides are known. The best known fatty acid amide is N-arachidonoylethanolamine (anandamide), a fatty acid amide found
in the human brain that binds to the cannabinoid receptors. We have a long interest in the enzymes of fatty acid amide biosynthesis.
We identified an enzyme that oxidizes the NAGs to the primary fatty acid amides and showed that inhibiting this enzyme led to the
cellular accumulation of the NAGs. We have characterized several insect N-acyltransferases (from
D. melanogaster, B. mori,
and T.
castaneum) that catalyze the acyl-CoA-dependent formation of fatty acid amides from an amine acyl-acceptor substrate. Knock-out
experiments in
D. melanogaster
validate our
in vitro
substrate specific studies demonstrating that one novel N-acyltransferases, arylalkyl
N-acyltransferase-like 2 (AANATL2), does catalyze the formation of N-acyldopamines
in vivo
. We developed a straightforward
platform technology to rapidly identify substrates for our panel of uncharacterized insect N-acyltransferases. Our application of this
technology leads to identification of an enzyme in
D. melanogaster
, agmatine N-acetyltransferase (AgmNAT), which catalyzes the
formation of N-acetylagmatine, a virtually unknown metabolite. We have determined the X-ray structure of AgmNAT. Our work on
AgmNAT hints at an unknown reaction in arginine metabolism and points to a novel class on fatty acid amides, the N-acylagmatine.
The presentation will also include our results on the kinetic and chemical mechanisms of the novel N-acyltransferases.
Recent Publications
1.
Dempsey D R et al. (2017) Structural and mechanistic analysis of Drosophila melanogaster agmatine N-acetyltransferase,
an enzyme that catalyzes the formation of N-acetylagmatine. Sci. Rep. 7(1):13432.
2.
Aboalroub A A et al. (2017) Acetyl group coordinated progression through the catalytic cycle of an arylalkylamine
N-acetyltransferase. PLoS One. 12(5):e0177270.
3.
Jeffries K A et al. (2016) Glycine N-acyltransferase-like 3 is responsible for long-chain N-acylglycine formation in N18TG2
cells. J. Lip. Res. 57(5):781-790.
4.
Dempsey D R, Carpenter A M, Rodriguez Ospina S and Merkler D J (2015) Probing the chemical mechanism and critical
regulatory amino acid residues in of Drosophila melanogaster arylalkylamine N-acyltransferase like 2. Insect Biochem. Mol.
Biol. 66:1-12.
5.
Dempsey D R et al. (2015) Mechanistic and structural analysis of a Drosophila melanogaster enzyme, arylalkylamine
N-acetyltransferase like 7, an enzyme that catalyzes the formation of N-acetylarylalkylamides and N-acetylhistamine.
Biochemistry. 54(16):2644-2658.
Biography
David J Merkler obtained a PhD in Biochemistry from Pennsylvania State University in 1985 and completed Postdoctoral Fellowships in Enzymology at Temple
University School of Medicine (1985-1987) and the Albert Einstein College of Medicine (1987-1989). His next position was as Senior Scientist at Unigene
Laboratories, Inc. involved in the
in vitro
production of a peptide hormone, calcitonin. In 1995, he moved back to academia as a Professor of Chemistry and
Biochemistry first at Duquesne University (1995-1999) and then the University of South Florida (1999-present). His laboratory has been interested in the fatty
amides: identification and characterization of the fatty acid amides (Lipidomics), identification and characterization of the enzymes of fatty acid amide biosynthesis
(Enzymology and Structural Biology), and changes in the fatty acid amidome after targeted enzyme knock-out (subtraction lipidomics).
merkler@usf.eduDavid J Merkler, J Biotechnol Biomater 2018, Volume 8
DOI: 10.4172/2155-952X-C2-091