The crystal structure of nitrophorin 2. A trifunctional antihemostatic protein from the saliva of Rhodnius prolixus

John F. Andersen, William "Bill" Montfort

Research output: Contribution to journalArticle

85 Citations (Scopus)

Abstract

Nitrophorin 2 (NP2) (also known as prolixin-S) is a salivary protein that transports nitric oxide, binds histamine, and acts as an anticoagulant during blood feeding by the insect Rhodnius prolixus. The 2.0-Å crystal structure of NP2 reveals an eight-stranded antiparallel β-barrel containing a ferric heme coordinated through His57, similar to the structures of NP1 and NP4. All four Rhodnius nitrophorins transport NO and sequester histamine through heme binding, but only NP2 acts as an anticoagulant. Here, we demonstrate that recombinant NP2, but not recombinant NP1 or NP4, is a potent anticoagulant; recombinant NP3 also displays minor activity. Comparison of the nitrophorin structures suggests that a surface region near the C terminus and the loops between β strands B-C and E-F is responsible for the anticoagulant activity. NP2 also displays larger NO association rates and smaller dissociation rates than NP1 and NP4, which may result from a more open and more hydrophobic distal pocket, allowing more rapid solvent reorganization on ligand binding. The NP2 protein core differs from NP1 and NP4 in that buried Glu53, which allows for larger NO release rates when deprotonated, hydrogen bonds to invariant Tyr81. Surprisingly, this tyrosine lies on the protein surface in NP1 and NP4.

Original languageEnglish (US)
Pages (from-to)30496-30503
Number of pages8
JournalJournal of Biological Chemistry
Volume275
Issue number39
StatePublished - Sep 29 2000

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Rhodnius
Saliva
Crystal structure
Anticoagulants
Proteins
Heme
Histamine
Salivary Proteins and Peptides
nitrophorin
Protein Transport
Tyrosine
Insects
Hydrogen
Hydrogen bonds
Nitric Oxide
Membrane Proteins
Blood
Association reactions
Ligands

ASJC Scopus subject areas

  • Biochemistry

Cite this

The crystal structure of nitrophorin 2. A trifunctional antihemostatic protein from the saliva of Rhodnius prolixus. / Andersen, John F.; Montfort, William "Bill".

In: Journal of Biological Chemistry, Vol. 275, No. 39, 29.09.2000, p. 30496-30503.

Research output: Contribution to journalArticle

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