While the three-dimensional structures of heme- and flavin-binding domains of the NOS isoforms have been determined, the structures of the holoenzymes remained elusive. Application of electron cryo-microscopy and structural modeling of the bovine endothelial nitric oxide synthase (eNOS) holoenzyme produced detailed models of the intact holoenzyme in the presence and absence of Ca(2+)/calmodulin (CaM). These models accommodate the cross-electron transfer from the reductase in one monomer to the heme in the opposite monomer. The heme domain acts as the anchoring dimeric structure for the entire enzyme molecule, while the FMN domain is activated by CaM to move flexibly to bridge the distance between the reductase and oxygenase domains. Our results indicate that the key regulatory role of CaM involves the stabilization of structural intermediates and precise positioning of the pivot for the FMN domain tethered shuttling motion to accommodate efficient and rapid electron transfer in the homodimer of eNOS.

• Klíčová slova
• Calmodulin, Electron cryomicroscopy, Electron transfer, Image processing, Nitric oxide synthase, Three-dimensional reconstruction,
• MeSH
• alosterická regulace MeSH
• flavinmononukleotid chemie   MeSH
• hem chemie   MeSH
• holoenzymy chemie   MeSH
• kalmodulin chemie   metabolismus   MeSH
• kinetika MeSH
• oxidace-redukce MeSH
• skot MeSH
• synthasa oxidu dusnatého, typ III chemie   metabolismus   MeSH
• terciární struktura proteinů MeSH
• transport elektronů MeSH
• vápník chemie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• flavinmononukleotid MeSH
• hem MeSH
• holoenzymy MeSH
• kalmodulin MeSH
• synthasa oxidu dusnatého, typ III MeSH
• vápník MeSH