Direct measurement by laser flash photolysis of intramolecular electron transfer in a two-domain construct of murine inducible nitric oxide synthase

Journal of the American Chemical Society
Changjian FengJohn H Enemark

Abstract

Intersubunit intramolecular electron transfer (IET) from FMN to heme is essential in the delivery of electrons required for O2 activation in the heme domain and the subsequent nitric oxide (NO) synthesis by NO synthase (NOS). Previous crystal structures and functional studies primarily concerned an enzyme conformation that serves as the input state for reduction of FMN by electrons from NADPH and FAD in the reductase domain. To favor formation of the output state for the subsequent IET from FMN to heme in the oxygenase domain, a novel truncated two-domain oxyFMN construct murine inducible nitric oxide synthase (iNOS), in which only the FMN and heme domains were present, was designed and expressed. The kinetics of the IET between the FMN and heme domains in this construct was directly determined using laser flash photolysis of CO dissociation in comparative studies on partially reduced oxyFMN and single domain heme oxygenase constructs.

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Citations

Oct 3, 2008·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Changjian FengDipak K Ghosh
Mar 13, 2012·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Wenbing LiChangjian Feng
Mar 18, 2009·Biochemistry·Robielyn P IlaganDennis J Stuehr
Aug 11, 2010·Journal of the American Chemical Society·Andrei V AstashkinChangjian Feng
May 2, 2009·Journal of the American Chemical Society·Joseph SempombeChangjian Feng
Jun 7, 2012·The Journal of Physical Chemistry. a·Andrei V AstashkinChangjian Feng
Aug 20, 2009·Dalton Transactions : an International Journal of Inorganic Chemistry·Changjian Feng, Gordon Tollin
Sep 5, 2013·Proceedings of the National Academy of Sciences of the United States of America·Brian C SmithMichael A Marletta
Jun 10, 2010·The Journal of Biological Chemistry·Jesús TejeroDennis J Stuehr
Oct 11, 2008·Biochimica Et Biophysica Acta·Donald E SprattJ Guy Guillemette
Dec 11, 2014·The FEBS Journal·Nicole G H LeferinkNigel S Scrutton
Oct 30, 2016·The Journal of Biological Chemistry·Mohammad Mahfuzul HaqueDennis J Stuehr

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