1X8N

1.08 A Crystal Structure Of Nitrophorin 4 From Rhodnius Prolixus Complexed With Nitric Oxide at pH 7.4

Summary for 1X8N

• Entry DOI: 10.2210/pdb1x8n/pdb
• Related: 1D2U 1KOI 1X8O 1X8P 1X8Q
• Descriptor: Nitrophorin 4, PROTOPORPHYRIN IX CONTAINING FE, NITRIC OXIDE, ... (4 entities in total)
• Functional Keywords: lipocalin; beta barrel; heme; nitric oxide, ligand binding protein
• Biological source: Rhodnius prolixus
• Cellular location: Secreted: Q94734
• Total number of polymer chains: 1
• Total formula weight: 20939.16

Authors

Kondrashov, D.A.,Roberts, S.A.,Weichsel, A.,Montfort, W.R. (deposition date: 2004-08-18, release date: 2004-10-05, Last modification date: 2023-08-23)

Primary citation

Kondrashov, D.A.,Roberts, S.A.,Weichsel, A.,Montfort, W.R.
Protein functional cycle viewed at atomic resolution: conformational change and mobility in nitrophorin 4 as a function of pH and NO binding
Biochemistry, 43:13637-13647, 2004

PubMed Abstract: The blood-sucking insect Rhodnius prolixus uses nitrophorin 4, a heme protein, to deliver nitric oxide (NO) to a victim, causing vasodilation and improved feeding. Binding of NO occurs at a ferric heme and is modulated by pH. NO binding at lower pH induces a large conformational change involving loops A-B and G-H that leads to distal pocket desolvation and protection of the nitrosyl heme complex. We have determined the crystal structures of Rhodnius nitrophorin 4 to ultrahigh resolution in four functional states: +/-NO at pH = 7.4 and +/-NO at pH = 5.6. The structure with NO at pH 7.4 (1.08 A) is newly determined while the other complexes have been modeled to resolutions much greater than previously reported (1.0-0.85 A). The ultrahigh resolution allowed us to resolve multiple conformers in binding-site loops, leading to a detailed description of the dynamics involved with storing NO in the insect salivary gland at low pH, and releasing NO in response to the increased pH of a victim's tissue. Strikingly, features for both the "open" and "closed" conformers exist under all conditions, suggesting that the flexible loops can transition with relative ease between conformational states. Yet, release of NO from rNP4 is much slower than found for other ferric heme proteins. The structures suggest that highly mobile loops can limit diffusion of diatomic molecules into and out of a protein cavity, a result with implications for the role of protein dynamics in function.

PubMed: 15504026
DOI: 10.1021/bi0483155

Experimental method

X-RAY DIFFRACTION (1.08 Å)