Analyse des Pheromonbindeprotein-Pheromon-Komplexes des Seidenspinners durch Elektrospray-Ionisierungs-Massenspektrometrie

Oldham, Neil J.; Krieger, Jürgen; Breer, Heinz; Fischedick, Annette; Hoskovec, Michal; Svatoš, Aleš

doi:10.1002/1521-3757(20001201)112:23<4521::aid-ange4521>3.0.co;2-9

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“…The amplified PCR fragments were purified and digested with MscI and XbaI before being cloned between the same restriction sites of the pET22b(+) vector. The recombinant pET22b(+)/ApolPBP2 and pET22b(+)/ApolPBP3 plasmids were transformed into E. coli BL21(DE3) and expression was performed following the protocols described for ApolPBP1 and the Bombyx mori PBP (Campanacci et al 2001;Oldham et al 2000) by growing the cells without induction at 28°C in LB medium supplemented with 50 lg/ll carbenicillin until A 600 reaches a value >2.5.…”

Section: Subcloning Of Pbps In Pet22b(+) and Bacterial Expressionmentioning

confidence: 99%

Species-specific pheromonal compounds induce distinct conformational changes of pheromone binding protein subtypes from Antheraea polyphemus

Mohl

Breer

Krieger

2002

Invertebrate Neuroscience

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We have investigated the structural features of three pheromone binding protein (PBP) subtypes from Antheraea polyphemus and monitored possible changes induced upon interaction with the Antheraea pheromonal compounds 4E,9Z-14:Ac [(E4,Z9)-tetradecadienyl-1-acetate], 6E,11Z-16:Ac [(E6,Z11)-hexadecadienyl-1-acetate], and 6E,11Z-16:Al [(E6,Z11)-hexadecadienal]. Circular dichroism and second derivative UV-difference spectroscopy data demonstrate that the structure of subtype PBP1 significantly changes upon binding of 4E,9Z-14:Ac. The related 6E,11Z-16:Ac was less effective and 6E,11Z-16:Al showed only a small effect. In contrast, in subtype PBP2 pronounced structural changes were only induced by the 6E,11Z-16:Al, and the subtype PBP3 did not show any considerable changes in response to the pheromonal compounds. The UV-spectroscopic data suggest that histidine residues are likely to be involved in the ligand-induced structural changes of the proteins, and this notion was confirmed by site-directed mutagenesis experiments. These results demonstrate that appropriate ligands induce structural changes in PBPs and provide evidence for ligand specificity of these proteins.

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Section: Subcloning Of Pbps In Pet22b(+) and Bacterial Expressionmentioning

confidence: 99%

Species-specific pheromonal compounds induce distinct conformational changes of pheromone binding protein subtypes from Antheraea polyphemus

Mohl

Breer

Krieger

2002

Invertebrate Neuroscience

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Analyse des Pheromonbindeprotein-Pheromon-Komplexes des Seidenspinners durch Elektrospray-Ionisierungs-Massenspektrometrie

Abstract: Garbenstraûe 30, 70593 Stuttgart (Deutschland) Dr. M. Hoskovec, Dr. A. Svatos Ï U Â stav organicke Â chemie a biochemie Akademie ve Ï d C Ï eske Â Republiky Flemingovo na Â m. 2, Prag 6-Dejvice, 16610 (Tschechische Republik)

Cited by 1 publication

References 20 publications

Species-specific pheromonal compounds induce distinct conformational changes of pheromone binding protein subtypes from Antheraea polyphemus

Species-specific pheromonal compounds induce distinct conformational changes of pheromone binding protein subtypes from Antheraea polyphemus

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