TITLE

Efficient expression of human soluble guanylate cyclase in Escherichia coli and its signaling-related interaction with nitric oxide

AUTHOR(S)
Fangfang Zhong; Hongyan Wang; Tianlei Ying; Zhong-Xian Huang; Xiangshi Tan
PUB. DATE
July 2010
SOURCE
Amino Acids;Jul2010, Vol. 39 Issue 2, p399
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Soluble guanylate cyclase (sGC), as a nitric oxide (NO) sensor, is a critical heme-containing enzyme in NO-signaling pathway of eukaryotes. Human sGC is a heterodimeric hemoprotein, composed of a α-subunit (690 AA) and a heme-binding β-subunit (619 AA). Upon NO binding, sGC catalyzes the conversion of guanosine 5′-triphosphate (GTP) to 3′,5′-cyclic guanosine monophosphate (cGMP). cGMP is a second messenger and initiates the nitric oxide signaling, triggering vasodilatation, smooth muscle relaxation, platelet aggregation, and neuronal transmission etc. The breakthrough of the bottle neck problem for sGC-mediated NO singling was made in this study. The recombinant human sGC β1 subunit (HsGCβ619) and its truncated N-terminal fragments (HsGCβ195 and HsGCβ384) were efficiently expressed in Escherichia coli and purified successfully in quantities. The three proteins in different forms (ferric, ferrous, NO-bound, CO-bound) were characterized by UV–vis and EPR spectroscopy. The homology structure model of the human sGC heme domain was constructed, and the mechanism for NO binding to sGC was proposed. The EPR spectra showed a characteristic of five-coordinated heme-nitrosyl species with triplet hyperfine splitting of NO. The interaction between NO and sGC was investigated and the schematic mechanism was proposed. This study provides new insights into the structure and NO-binding of human sGC. Furthermore, the efficient expression system of E. coli will be beneficial to the further studies on structure and activation mechanism of human sGC.
ACCESSION #
51652523

 

Related Articles

  • New insight into the functioning of nitric oxide-receptive guanylyl cyclase: physiological and pharmacological implications. John Garthwaite // Molecular & Cellular Biochemistry;Jan2010, Vol. 334 Issue 1/2, p221 

    Abstract  The cellular counterpart of the “soluble” guanylyl cyclase found in tissue homogenates over 30 years ago is now recognized as the physiological receptor for nitric oxide (NO). The ligand-binding site is a prosthetic haem group that, when occupied by NO, induces a...

  • The alpha1 isoform of soluble guanylate cyclase regulates cardiac contractility but is not required for ischemic preconditioning. Sips, Patrick; Brouckaert, Peter; Ichinose, Fumito // Basic Research in Cardiology;Jul2011, Vol. 106 Issue 4, p635 

    Nitric oxide (NO)-dependent soluble guanylate cyclase (sGC) activation is an important component of cardiac signal transduction pathways, including the cardioprotective signaling cascade induced by ischemic preconditioning (IPC). The sGCα subunit, which binds to the common sGCβ1 subunit,...

  • Ghrelin stimulates proliferation of human osteoblastic TE85 cells via NO/cGMP signaling pathway. Deng-Hu Wang; Yun-Sheng Hu; Jun-Jie Du; Yun-Yu Hu; Wei-De Zhong; Wei-Jun Qin // Endocrine (1355008X);Feb2009, Vol. 35 Issue 1, p112 

    Abstract  Ghrelin regulates bone formation and osteoblast proliferation, but the detailed signaling pathway for its action on osteoblasts remains unclear. In human osteoblastic TE85 cells, we observed the effects and intracellular signaling pathway of ghrelin on cell proliferation using...

  • A novel insight into the heme and NO/CO binding mechanism of the alpha subunit of human soluble guanylate cyclase. Zhong, Fangfang; Pan, Jie; Liu, Xiaoxiao; Wang, Hongyan; Ying, Tianlei; Su, Jihu; Huang, Zhong-Xian; Tan, Xiangshi // Journal of Biological Inorganic Chemistry;Dec2011, Vol. 16 Issue 8, p1227 

    Human soluble guanylate cyclase (sGC), a critical heme-containing enzyme in the NO-signaling pathway of eukaryotes, is an αβ heterodimeric hemoprotein. Upon the binding of NO to the heme, sGC catalyzes the conversion of GTP to cyclic GMP, playing a crucial role in many physiological...

  • Crystal Structures of the Catalytic Domain of Human Soluble Guanylate Cyclase. Allerston, Charles K.; von Delft, Frank; Gileadi, Opher // PLoS ONE;Mar2013, Vol. 8 Issue 3, p1 

    Soluble guanylate cyclase (sGC) catalyses the synthesis of cyclic GMP in response to nitric oxide. The enzyme is a heterodimer of homologous α and β subunits, each of which is composed of multiple domains. We present here crystal structures of a heterodimer of the catalytic domains of the...

  • Elevation of Soluble Guanylate Cyclase Suppresses Proliferation and Survival of Human Breast Cancer Cells. Wen, Hui-Chin; Chuu, Chih-Pin; Chen, Chen-Yu; Shiah, Shine-Gwo; Kung, Hsing-Jien; King, Kuang-Liang; Su, Liang-Chen; Chang, Shi-Chuan; Chang, Chung-Ho // PLoS ONE;Apr2015, Vol. 10 Issue 4, p1 

    Nitric oxide (NO) is an essential signaling molecule in biological systems. Soluble guanylate cyclase (sGC), composing of α1 and β1 subunit, is the receptor for NO. Using radioimmunoassay, we discovered that activation of sGC by treatment with bradykinin or sodium nitroprusside (SNP) is...

  • Cyclic Stretch Induces Inducible Nitric Oxide Synthase and Soluble Guanylate Cyclase in Pulmonary Artery Smooth Muscle Cells. Shah, Monica R.; Wedgwood, Stephen; Czech, Lyubov; Kim, Gina A.; Lakshminrusimha, Satyan; Schumacker, Paul T.; Steinhorn, Robin H.; Farrow, Kathryn N. // International Journal of Molecular Sciences;Feb2013, Vol. 14 Issue 2, p4334 

    In the pulmonary vasculature, mechanical forces such as cyclic stretch induce changes in vascular signaling, tone and remodeling. Nitric oxide is a potent regulator of soluble guanylate cyclase (sGC), which drives cGMP production, causing vasorelaxation. Pulmonary artery smooth muscle cells...

  • Dual effect of nitric oxide on ATP-sensitive K+ channels in rat pancreatic β cells. Sunouchi, Takaaki; Suzuki, Kimiaki; Nakayama, Koichi; Ishikawa, Tomohisa // Pflugers Archiv European Journal of Physiology;Jun2008, Vol. 456 Issue 3, p573 

    We have previously shown that NO has stimulatory and inhibitory effects on insulin secretion at low and high concentrations, respectively. The present study investigated effects of NO on KATP channels of rat β cells by patch clamp analysis to elucidate the mechanism for the dual effect. NOC7...

  • Supersensitivity of soluble guanylyl cyclase mediates enhanced nitrovasodilator sensitivity in eNOS knockout mice.  // Pflugers Archiv European Journal of Physiology;Apr2000 Supplement, Vol. 439, pR423 

    An abstract of the study "Supersensitivity of soluble guanylyl cyclase mediates enhanced nitrovasodilator sensitivity in eNOS knockout mice" is presented. It states that endogenous nitric oxide (NO) weakens the nitrovasodilator-induced vasodilation. Results of the study reveals that chronic...

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics