Oligomerization of nitrophorins.
Rhodnius prolixus is a blood-sucking insect that uses a mixture of nitrophorin (NP) proteins to deliver nitric oxide (NO) from the insect saliva to the hosts via a ferric heme coordinated to the protein, causing vasodilatation and anticoagulation to support their feeding. R. prolixus NPs 1-4 are very similar proteins ( approximately 20 kDa) with different NO affinities for stepwise NO release triggered by pH increase and histamine binding in hosts. Ultra-high-resolution X-ray structures of native and mutant NPs and their kinetic analysis already have revealed the fundamental steps of NO binding and release. In this study, we found that NPs can exist in multiple oligomerization states at higher concentrations. The oligomers are characterized by a combination of multiple biophysical methods. The intrinsic features of the oligomerization revealed here led us to propose that this intensive, moderately pH- and ligand-dependent oligomerization of NPs has physiological implications in the facilitation of the efficient storage and release of the highly reactive NO in the insect saliva and the victim, respectively.
Department of Pharmacology and Toxicology, University of Arizona, Tucson, 85721, USA. ambrus@pharmacy.arizona.edu
Attila
A
Ambrus
Kenneth
K
Friedrich
Arpád
A
Somogyi
D000818:Animals; D001511:Bedbugs; D002853:Chromatography, Liquid; D006420:Hemeproteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D013058:Mass Spectrometry; D009154:Mutation; D017942:Nitric Acid / Q000737:chemistry; D009841:Oligonucleotides / Q000737:chemistry*; D012239:Rhodnius; D012471:Salivary Proteins and Peptides / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D006420:Hemeproteins; D008024:Ligands; D009841:Oligonucleotides; D012471:Salivary Proteins and Peptides; C090560:nitrophorin; D017942:Nitric Acid