Duality monomer-dimer of the pheromone-binding protein from Bombyx mori.
The analysis of a recombinant pheromone-binding protein from the silkworm moth, Bombyx mori, by native gel electrophoresis with Coomassie staining showed one single band with a molecular mass consistent with a monomer. A slow migrating band, detected in the recombinant and native samples by a polyclonal antibody, was indistinguishable from the monomer in the mass spectrum fragmentation pattern and chromatographic behavior. Flow injection analyses of the protein by mass spectrometry in the negative mode showed fragments of a dimer. The dimeric form was also supported by estimation of the molecular mass by gel filtration at basic pH. A cross-linked dimer coeluted with the noncovalent dimer on a gel filtration column. The molecular mass of the protein changed in a pH-dependent way with a dramatic transition from dimer to monomer between pH 6 and 4.5. A low pH induced not only dissociation of the dimer, but also a conformational change in the protein. In marked contrast to denaturation with guanidinium chloride, the emission maxima of tryptophan was not significantly changed at low pH. BmPBP is thus a dimer at slightly acid, neutral, and basic pH, which dissociates and then undergoes conformational change at low pH.
Leal
W S
WS
Laboratory of Chemical Prospecting, National Institute of Sericultural and Entomological Science, Tsukuba, Japan. leal@nises.affrc.go.jp
D000818:Animals; D015153:Blotting, Western; D012831:Bombyx / Q000737:chemistry*; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry; D019281:Dimerization; D019476:Insect Proteins / Q000737:chemistry* / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins; D013058:Mass Spectrometry; D008970:Molecular Weight; D013050:Spectrometry, Fluorescence
D002352:Carrier Proteins; D003432:Cross-Linking Reagents; D019476:Insect Proteins; D036341:Intercellular Signaling Peptides and Proteins; C403094:pheromone-binding protein, Bombyx