Versatile Regulation of Neuronal Nitric Oxide Synthase by Specific Regions of Its C-Terminal Tail
Abstract
The C-terminal tail (CT) of neuronal nitric oxide synthase (nNOS) is a regulatory element that suppresses nNOS activities in the absence of bound calmodulin (CaM). A crystal structure of the nNOS reductase domain (nNOSr) (Garcin, E. D., Bruns, C. M., Lloyd, S. J., Hosfield, D. J., Tiso, M., Gachhui, R., Stuehr, D. J., Tainer, J. A., and Getzoff, E. D. (2004) J. Biol. Chem. 279, 37918-37927) revealed how the first half of the CT interacts with nNOSr and thus provided a template for detailed studies. We generated truncation mutants in nNOS and nNOSr to test the importance of 3 different regions of the CT. Eliminating the terminal half of the CT (all residues from Ile1413 to Ser1429), which is invisible in the crystal structure, had almost no impact on NADP+ release, flavin reduction, flavin autoxidation, heme reduction, reductase activity, or NO synthesis activity, but did prevent an increase in FMN shielding that normally occurs in response to NADPH binding. Additional removal of the CT alpha-helix (residues 1401 to 1412) significantly increased the NADP+ release rate, flavin autoxidation, and NADPH oxidase activity, and caused hyper-deshielding of the FMN cofactor. These effects were associated with increased reductase activity and slightly diminished heme reduction and NO synthesis. Further removal of residues downstream from Gly1396 (a full CT truncation) amplified the aforementioned effects and in addition altered NADP+ interaction with FAD, relieved the kinetic suppression on flavin reduction, and further diminished heme reduction and NO synthesis. Our results reveal that the CT exerts both multifaceted and regiospecific effects on catalytic activities and related behaviors, and thus provide new insights into mechanisms that regulate nNOS catalysis.
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Deoxygenated Ngb can reduce nitrite to NO.
“It was recently demonstrated that, as observed with other globins, deoxygenated Ngb can also reduce nitrite to NO (932, 945).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [945]
Cites
PROCHECK: a program to check the stereochemical quality of protein structures
Laskowski, R. A., MacArthur, M. W., Moss, D. S., & Thornton, J. M. (1993) · Journal of Applied Crystallography · doi:10.1107/s0021889892009944 · Cited by 24,581 · see citing works
SWISS‐MODEL and the Swiss‐Pdb Viewer: An environment for comparative protein modeling
Guex, N., & Peitsch, M. (1997) · Electrophoresis · doi:10.1002/elps.1150181505 · Cited by 11,589 · see citing works
SWISS-MODEL: an automated protein homology-modeling server
Schwede, T. F. (2003) · Nucleic Acids Research · doi:10.1093/nar/gkg520 · Cited by 5,592 · see citing works
Nitric oxide synthases: structure, function and inhibition
ALDERTON, W. K., COOPER, C. E., & KNOWLES, R. G. (2001) · Biochemical Journal · doi:10.1042/bj3570593 · Cited by 4,110 · see citing works
Nitric oxide synthases: structure, function and inhibition
ALDERTON, W. K., Cooper, C. E., & Knowles, R. G. (2001) · Biochemical Journal · doi:10.1042/0264-6021:3570593 · Cited by 4,106 · see citing works
Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase
Bredt, D. S., Hwang, P. M., Glatt, C. E., Lowenstein, C. J., Reed, R. R., & Snyder, S. H. (1991) · Nature · doi:10.1038/351714a0 · Cited by 2,354 · see citing works
Nitric oxide synthases: which, where, how, and why?
Michel, T. M., & Féron, O. (1997) · Journal of Clinical Investigation · doi:10.1172/jci119750 · Cited by 1,018 · see citing works
Mammalian nitric oxide synthases
Stuehr, D. J. (1999) · Biochimica et Biophysica Acta (BBA) - Bioenergetics · doi:10.1016/s0005-2728(99)00016-x · Cited by 996 · see citing works
Self-citationNitric oxide in health and disease of the respiratory system
Ricciardolo, F. L. M., Sterk, P. J., Gaston, B., & Folkerts, G. (2004) · Physiological Reviews · doi:10.1152/physrev.00034.2003 · Cited by 903 · see citing works
Wang, M., Roberts, D. L., Paschke, R., Shea, T. M., Masters, B. S. S., & Kim, J.-J. P. (1997) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.94.16.8411 · Cited by 744 · see citing works
Nitric oxide synthases reveal a role for calmodulin in controlling electron transfer.
Abu‐Soud, H. M., & Stuehr, D. J. (1993) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.90.22.10769 · Cited by 410 · see citing works
Self-citationStuehr, D. J., & Ikeda‐Saito, M. (1992) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)36718-3 · Cited by 348 · see citing works
Self-citationNitric oxide: discovery and impact on clinical medicine
Moncada, S. (1999) · Journal of the Royal Society of Medicine · doi:10.1177/014107689909200402 · Cited by 337 · see citing works
Structure–function studies on nitric oxide synthases
LI, H., & POULOS, T. (2005) · Journal of Inorganic Biochemistry · doi:10.1016/j.jinorgbio.2004.10.016 · Cited by 302 · see citing works
Structural Basis for Isozyme-specific Regulation of Electron Transfer in Nitric-oxide Synthase
Garcin, E. D., Bruns, C. M., Lloyd, S. J., Hosfield, D. J., Tiso, M., Gachhui, R., Stuehr, D. J., Tainer, J. A., & Getzoff, E. D. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m406204200 · Cited by 285 · see citing works
Self-citationAbu‐Soud, H. M., Yoho, L. L., & Stuehr, D. J. (1994) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)31597-7 · Cited by 249 · see citing works
Self-citationAn Autoinhibitory Control Element Defines Calcium-regulated Isoforms of Nitric Oxide Synthase
Salerno, J. C., Harris, D. E., Irizarry, K., Patel, B., Morales, A. J., Smith, S. M., Pavel Martasek, E., Roman, L. J., Masters, B. S. S., Jones, C. L., et al. (1997) · Journal of Biological Chemistry · doi:10.1074/jbc.272.47.29769 · Cited by 248 · see citing works
NADPH-Cytochrome P450 Oxidoreductase
Hubbard, P. A., Shen, A. L., Paschke, R., Kasper, C. B., & Kim, J. P. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m101731200 · Cited by 217 · see citing works
Karplus, P. A., Deng, Z., Aliverti, A., Zanetti, G., Arakaki, A. K., Ottado, J., Orellano, E. G., Calcaterra, N. B., Ceccarelli, E. A., & Carrillo, N. J. (1999) · Nature Structural Biology · doi:10.1038/12307 · Cited by 200 · see citing works
Masters, B. S. S., McMillan, K., Sheta, E. A., Nishimura, J. S., Roman, L. J., & Pavel Martasek, E. (1996) · The FASEB Journal · doi:10.1096/fasebj.10.5.8621055 · Cited by 180 · see citing works
Tetrahydrobiopterin Radical Enzymology
Wei, C.-C., Crane, B. R., & Stuehr, D. J. (2003) · Chemical Reviews · doi:10.1021/cr0204350 · Cited by 175 · see citing works
Self-citationGachhui, R., Presta, A., Bentley, D. F., Abu‐Soud, H. M., McArthur, R., Brudvig, G. W., Ghosh, D. K., & Stuehr, D. J. (1996) · Journal of Biological Chemistry · doi:10.1074/jbc.271.34.20594 · Cited by 142 · see citing works
Self-citationCrystal Structure of the FAD/NADPH-binding Domain of Rat Neuronal Nitric-oxide Synthase
Zhang, J., Martásek, P., Paschke, R., et al. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m105503200 · Cited by 136 · see citing works
Neuronal Nitric Oxide Synthase
Gerber, N. C., & de Montellano, P. R. O. (1995) · Journal of Biological Chemistry · doi:10.1074/jbc.270.30.17791 · Cited by 133 · see citing works
Daff, S. N., Sagami, I., & Shimizu, T. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.43.30589 · Cited by 126 · see citing works
Potentiometric Analysis of the Flavin Cofactors of Neuronal Nitric Oxide Synthase
Noble, M. A., Munro, A. W., Rivers, S. L., Robledo, L., Daff, S. N., Yellowlees, L. J., Shimizu, T., Sagami, I., Guillemette, J. G., & Chapman, S. K. (1999) · Biochemistry · doi:10.1021/bi992150w · Cited by 117 · see citing works
Novel features of nitric oxide, endothelial nitric oxide synthase, and atherosclerosis
Ignarro, L. J., & Claudio Napoli, A. (2005) · Current Diabetes Reports · doi:10.1007/s11892-005-0062-8 · Cited by 116 · see citing works
Roman, L. J., Pavel Martasek, E., Miller, R. T., Harris, D. E., de la Garza, M. A., Shea, T. M., Kim, J. J., & Masters, B. S. S. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m004766200 · Cited by 115 · see citing works
Adak, S., Santolini, J., Tikunova, S. B., Wang, Q., Johnson, J., & Stuehr, D. J. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m006857200 · Cited by 115 · see citing works
Self-citationCRAIG, D. H., Chapman, S. K., & Daff, S. N. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m203118200 · Cited by 99 · see citing works
Siddhanta, U., Wu, C., Abu‐Soud, H. M., Zhang, J., Ghosh, D. K., & Stuehr, D. J. (1996) · Journal of Biological Chemistry · doi:10.1074/jbc.271.13.7309 · Cited by 95 · see citing works
Self-citationPiubelli, L., Aliverti, A., Arakaki, A. K., et al. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.275.14.10472 · Cited by 94 · see citing works
Roman, L. J., & Masters, B. S. S. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m603671200 · Cited by 93 · see citing works
Role of Reductase Domain Cluster 1 Acidic Residues in Neuronal Nitric-oxide Synthase
Adak, S., Ghosh, S., Abu‐Soud, H. M., & Stuehr, D. J. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.32.22313 · Cited by 89 · see citing works
Self-citationMatsuda, H., & Iyanagi, T. (1999) · Biochimica et Biophysica Acta (BBA) - General Subjects · doi:10.1016/s0304-4165(99)00193-2 · Cited by 83 · see citing works
Lane, P. B., & Gross, S. S. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m200258200 · Cited by 80 · see citing works
Wu, C., Zhang, J., Abu‐Soud, H. M., et al. (1996) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.1996.0763 · Cited by 76 · see citing works
Gachhui, R., Ghosh, D. K., Wu, C., et al. (1997) · Biochemistry · doi:10.1021/bi970331x · Cited by 75 · see citing works
Hurshman, A. R., & Marletta, M. A. (2002) · Biochemistry · doi:10.1021/bi012002h · Cited by 71 · see citing works
Roman, L. J., Miller, R. T., de la Garza, M. A., Kim, J. J., & Masters, B. S. S. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m002449200 · Cited by 69 · see citing works
Neuronal Nitric-oxide Synthase Interaction with Calmodulin-Troponin C Chimeras
Gachhui, R., Abu‐Soud, H. M., Ghoshà, D. K., Presta, A., Blazing, M. A., Mayer, B., George, S. E., & Stuehr, D. J. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.10.5451 · Cited by 69 · see citing works
Self-citationFeng, C., Tollin, G., Holliday, M. A., Thomas, C., Salerno, J. C., Enemark, J. H., & Ghosh, D. K. (2006) · Biochemistry · doi:10.1021/bi060223n · Cited by 68 · see citing works
Montgomery, H. J., Romanov, V., & Guillemette, J. G. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.275.7.5052 · Cited by 66 · see citing works
Knight, K., & Scrutton, N. S. (2002) · Biochemical Journal · doi:10.1042/bj20020667 · Cited by 65 · see citing works
Chen, P., & Wu, K. K. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.275.17.13155 · Cited by 64 · see citing works
Surface Charge Interactions of the FMN Module Govern Catalysis by Nitric-oxide Synthase
Panda, K., Haque, M. M., Garcin, E. D., Durra, D., Getzoff, E. D., & Stuehr, D. J. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m606129200 · Cited by 62 · see citing works
Self-citationKnudsen, G. M., Nishida, C. R., Mooney, S. D., et al. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m303267200 · Cited by 60 · see citing works
C-Terminal Tyrosine of Ferredoxin−NADP+ Reductase in Hydride Transfer Processes with NAD(P)+/H
Tejero, J., Pérez‐Dorado, I., Maya, C., et al. (2005) · Biochemistry · doi:10.1021/bi051278c · Cited by 54 · see citing works
Self-citationConformational and Thermodynamic Control of Electron Transfer in Neuronal Nitric Oxide Synthase
Dunford, A. J., Rigby, S. E. J., Hay, S., Munro, A. W., & Scrutton, N. S. (2007) · Biochemistry · doi:10.1021/bi7001339 · Cited by 54 · see citing works
Konas, D. W., Zhu, K., Sharma, M., Aulak, K. S., Brudvig, G. W., & Stuehr, D. J. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m400872200 · Cited by 51 · see citing works
Self-citationControl of Electron Transfer in Nitric-oxide Synthases
Nishida, C. R., & de Montellano, P. R. O. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m101548200 · Cited by 51 · see citing works
Grunau, A., Paine, M. J. I., Ladbury, J. E., et al. (2006) · Biochemistry · doi:10.1021/bi052115r · Cited by 45 · see citing works
Adak, S., Sharma, M., Meade, A. L., & Stuehr, D. J. (2002) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.192283399 · Cited by 44 · see citing works
Self-citationCalcium-Binding Sites of Calmodulin and Electron Transfer by Inducible Nitric Oxide Synthase
Gribovskaja, I., Brownlow, K. C., Dennis, S. J., et al. (2005) · Biochemistry · doi:10.1021/bi0474517 · Cited by 42 · see citing works
Tiso, M., Konas, D. W., Panda, K., et al. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m507775200 · Cited by 42 · see citing works
Self-citationControl of electron transfer in neuronal NO synthase
Daff, S. N., Noble, M., CRAIG, D. H., et al. (2001) · Biochemical Society Transactions · doi:10.1042/0300-5127:0290147 · Cited by 36 · see citing works
Control of electron transfer in neuronal NO synthase
Daff, S. N., Noble, M., CRAIG, D. H., et al. (2001) · Biochemical Society Transactions · doi:10.1042/bst0290147 · Cited by 36 · see citing works
Chen, P., & Wu, K. K. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m305469200 · Cited by 33 · see citing works
Garnaud, P. E., Koetsier, M. J., Ost, T. W. B., & Daff, S. N. (2004) · Biochemistry · doi:10.1021/bi049312v · Cited by 30 · see citing works
Grunau, A., Geraki, K., Grossmann, J. G., et al. (2007) · Biochemistry · doi:10.1021/bi700596s · Cited by 29 · see citing works
Dunford, A. J., Marshall, K. R., Munro, A. W., et al. (2004) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.2004.04185.x · Cited by 28 · see citing works
Interactions between the Isolated Oxygenase and Reductase Domains of Neuronal Nitric-oxide Synthase
Rozhkova, E. A., Fujimoto, N., Sagami, I., et al. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m200642200 · Cited by 26 · see citing works
A Conserved Aspartate (Asp-1393) Regulates NADPH Reduction of Neuronal Nitric-oxide Synthase
Panda, K., Adak, S., Konas, D. W., Sharma, M., & Stuehr, D. J. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m310391200 · Cited by 26 · see citing works
Self-citationWei, C., Wang, Z., & Stuehr, D. J. (2002) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(02)54026-x · Cited by 9 · see citing works
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Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation
Tejero, J., Shiva, S., & Gladwin, M. T. (2019) · Physiological Reviews · doi:10.1152/physrev.00036.2017 · Cited by 541 · see citing works
Self-citationNO synthase: Structures and mechanisms
Daff, S. (2010) · Nitric Oxide · doi:10.1016/j.niox.2010.03.001 · Cited by 251 · see citing works
- Authors
- Tiso, M., Tejero, J., Panda, K., Aulak, K. S., & Stuehr, D. J.
- Affiliations
- Cleveland State University (US)
- Corresponding author
- Tiso, Mauro
- Stuehr, Dennis J.
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2007-11-20
- Volume
- 46
- Issue
- 50
- Pages
- 14418-14428
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W1969901042
- Cited by
- 34
- References
- 64
- Citations by year
- 2026: 2 · 2018: 5 · 2016: 3 · 2014: 4 · 2013: 4 · 2012: 4
- FWCI
- 2.45
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Heart, Lung, and Blood Institute (HL76491, P01HL076491)
- National Institute of General Medical Sciences (GM51491, R01GM051491)
- Topics
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Hemoglobin structure and function
- Physical Sciences › Chemistry › Inorganic Chemistry › Metal-Catalyzed Oxygenation Mechanisms
- Keywords
- neuronal nitric oxide synthase, C-terminal tail, calmodulin, enzyme regulation, heme reduction, NADPH oxidase activity, truncation mutants, catalytic mechanism
- MeSH terms
- Amino Acid Sequence, Animals, Computer Simulation, Cytochrome Reductases, Flavin-Adenine Dinucleotide, Flavin Mononucleotide, Heme, Kinetics, Molecular Sequence Data, Mutation, NADP, Oxidation-Reduction, Protein Binding, Protein Structure, Secondary, Protein Structure, Tertiary, Rats, Nitric Oxide Synthase Type I
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