Lipoic acid is an essential sulfur-containing cofacor that is found in several multienzyme complexes that are involved in energy metabolism. In its functional form it is covaleritly attached to the epsilon amino group of a specific lysine residue on a designated lipoyl carrying protein. The detailed manner in which it functions in these complexes has been known for quite some time, and is found in almost every beginning biochemistry textbook. By contrast, only within the last few years has a clear understanding of its biosynthesis begun to emerge. Lipoic acid is biosynthesized in its cofactor form rather than its free acid form, requiring two proteins that are dedicated to the transformation. The first, octanoyl transferase (LipB) catalyzes the transfer of the eight-carbon fatty acyl chain from octanoyl-ACP to one of the several lipoyl carrying proteins in the cell. The second protein, lipoyl synthase (LipA) catalyzes the insertion of two sulfur atoms at positions 6 and 8 of the fatty acyl group, affording the lipoyl cofactor. LipA is a member of radical SAM superfamily of enzymes, which use a [4Fe-4S] and S-adenosylmethionine to generate high-energy radicals that are intermediates in each reaction. The long-term objective of this proposal is to understand at the detailed molecular level how each of these enzymes work. Particular focus will be directed at LipA, to understand exactly from where the sulfur atom that is inserted into the substrate is derived. Efforts will involve bacterial genetics, transient state kinetics, and spectroscopy (UV-vis, EPR, M""""""""ssbauer, ENDOR, and FT-ICR mass spectrometry). Aside from the involvement of lipoic acid as a cofactor in enzyme complexes of energy metabolism it is known to modulate glucose metabolism in patients with type II diabetes and to serve as a general cellular antioxidant, among many other things. There is significant evidence that lipoic acid can be endogenously synthesized in mammalian cells, and that the inhibition of this pathway compromises cellular function and leads to death.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM063847-09
Application #
7778833
Study Section
Macromolecular Structure and Function A Study Section (MSFA)
Program Officer
Jones, Warren
Project Start
2002-04-01
Project End
2012-02-28
Budget Start
2010-03-01
Budget End
2012-02-28
Support Year
9
Fiscal Year
2010
Total Cost
$249,192
Indirect Cost
Name
Pennsylvania State University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
003403953
City
University Park
State
PA
Country
United States
Zip Code
16802
McLaughlin, Martin I; Lanz, Nicholas D; Goldman, Peter J et al. (2016) Crystallographic snapshots of sulfur insertion by lipoyl synthase. Proc Natl Acad Sci U S A 113:9446-50
Lanz, Nicholas D; Lee, Kyung-Hoon; Horstmann, Abigail K et al. (2016) Characterization of Lipoyl Synthase from Mycobacterium tuberculosis. Biochemistry 55:1372-83
Lanz, Nicholas D; Booker, Squire J (2015) Auxiliary iron-sulfur cofactors in radical SAM enzymes. Biochim Biophys Acta 1853:1316-34
Pandelia, Maria-Eirini; Lanz, Nicholas D; Booker, Squire J et al. (2015) Mössbauer spectroscopy of Fe/S proteins. Biochim Biophys Acta 1853:1395-405
Lanz, Nicholas D; Rectenwald, Justin M; Wang, Bo et al. (2015) Characterization of a Radical Intermediate in Lipoyl Cofactor Biosynthesis. J Am Chem Soc 137:13216-9
Lanz, Nicholas D; Pandelia, Maria-Eirini; Kakar, Elizabeth S et al. (2014) Evidence for a catalytically and kinetically competent enzyme-substrate cross-linked intermediate in catalysis by lipoyl synthase. Biochemistry 53:4557-72
Goldman, Peter J; Grove, Tyler L; Sites, Lauren A et al. (2013) X-ray structure of an AdoMet radical activase reveals an anaerobic solution for formylglycine posttranslational modification. Proc Natl Acad Sci U S A 110:8519-24
Landgraf, Bradley J; Arcinas, Arthur J; Lee, Kyung-Hoon et al. (2013) Identification of an intermediate methyl carrier in the radical S-adenosylmethionine methylthiotransferases RimO and MiaB. J Am Chem Soc 135:15404-15416
Goldman, Peter J; Grove, Tyler L; Booker, Squire J et al. (2013) X-ray analysis of butirosin biosynthetic enzyme BtrN redefines structural motifs for AdoMet radical chemistry. Proc Natl Acad Sci U S A 110:15949-54
Grove, Tyler L; Ahlum, Jessica H; Qin, Rosie M et al. (2013) Further characterization of Cys-type and Ser-type anaerobic sulfatase maturating enzymes suggests a commonality in the mechanism of catalysis. Biochemistry 52:2874-87

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