Structural Studies of a Complex Between Endothelial Nitric Oxide Synthase and Calmodulin at Physiological Calcium Concentration
Abstract
The small acidic protein calmodulin (CaM) serves as a Ca 2+ sensor and control element for many enzymes including nitric oxide synthase (NOS) enzymes that play major roles in key physiological and pathological processes. CaM binding causes a conformational change in NOS to allow for the electron transfer between the reductase and oxygenase domains through a process that is thought to be highly dynamic. In this report, NMR spectroscopy was used to determine the solution structure of the endothelial NOS (eNOS) peptide in complex with CaM at the lowest Ca 2+ concentration (225 nM) required for CaM to bind to eNOS and corresponds to a physiological elevated Ca 2+ level found in mammalian cells. Under these conditions, the CaM–eNOS complex has a Ca 2+ -replete C-terminal lobe bound to the eNOS peptide and a Ca 2+ free N-terminal lobe loosely associated with the eNOS peptide. With increasing Ca 2+ concentration, the binding of Ca 2+ by the N-lobe of CaM results in a stronger interaction with the C-terminal region of the eNOS peptide and increased α-helical structure of the peptide that may be part of the mechanism resulting in electron transfer from the FMN to the heme in the oxygenase domain of the enzyme. Surface plasmon resonance studies performed under the same conditions show Ca 2+ concentration-dependent binding kinetics were consistent with the NMR structural results. This investigation shows that structural studies performed under more physiological relevant conditions provide information on subtle changes in structure that may not be apparent when experiments are performed in excess Ca 2+ concentrations.
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HO-mediated heme degradation generates ferrous iron (Fe2+) and CO.
“This process generates ferrous iron (Fe2+) and CO, which are well-known toxic species (122, 739, 795, 959), but CO can also be, in low concentrations, a signaling molecule (713, 739, 805, 807).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [739]
CO can be, in low concentrations, a signaling molecule.
“but CO can also be, in low concentrations, a signaling molecule (713, 739, 805, 807).”
From Sources of Vascular Nitric Oxide and Reactive Oxygen Species and Their Regulation (2019) · cites this paper as [739]
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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
- Authors
- Piazza, M., Dieckmann, T., & Guillemette, J. G.
- Affiliations
- University of Waterloo (CA)
- Corresponding author
- Dieckmann, Thorsten
- Guillemette, J. Guy
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2016-10-12
- Volume
- 55
- Issue
- 42
- Pages
- 5962-5971
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W2529811016
- Cited by
- 23
- References
- 58
- Citations by year
- 2024: 2 · 2023: 7 · 2022: 1 · 2021: 3 · 2020: 1 · 2019: 3 · 2018: 6
- FWCI
- 1.56
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- UWSpace (University of Waterloo) (submittedVersion)
- Funders
- Natural Sciences and Engineering Research Council of Canada (183521, 326911)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Hemoglobin structure and function
- Keywords
- endothelial nitric oxide synthase, calmodulin, protein-protein complex, calcium binding, solution structure, electron transfer, enzyme conformational change, calcium sensor, binding kinetics
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