Activation mechanism of recombinant Der p 3 allergen zymogen: contribution of cysteine protease Der p 1 and effect of propeptide glycosylation.
The trypsin-like protease Der p 3, a major allergen of the house dust mite Dermatophagoides pteronyssinus, is synthesized as a zymogen, termed proDer p 3. No recombinant source of Der p 3 has been described yet, and the zymogen maturation mechanism remains to be elucidated. The Der p 3 zymogen was produced in Pichia pastoris. We demonstrated that the recombinant zymogen is glycosylated at the level of its propeptide. We showed that the activation mechanism of proDer p 3 is intermolecular and is mediated by the house dust mite cysteine protease Der p 1. The primary structure of the proDer p 3 propeptide is associated with a unique zymogen activation mechanism, which is different from those described for the trypsin-like family and relies on the house dust mite papain-like protease Der p 1. This is the first report of a recombinant source of Der p 3, with the same enzymatic activity as the natural enzyme and trypsin. Glycosylation of the propeptide was found to decrease the rate of maturation. Finally, we showed that recombinant Der p 3 is inhibited by the free modified prosequence T(P1)R.
Dumez
Marie-Eve
ME
Centre for Protein Engineering, Centre de Recherches du Cyclotron, Université de Liège, Liège 4000, Belgium.
Teller
Nathalie
N
Mercier
Frédéric
F
Tanaka
Tetsuya
T
Vandenberghe
Isabel
I
Vandenbranden
Michel
M
Devreese
Bart
B
Luxen
André
A
Frère
Jean-Marie
JM
Matagne
André
A
Jacquet
Alain
A
Galleni
Moreno
M
Chevigné
Andy
A
D000595:Amino Acid Sequence; D000818:Animals; D039741:Antigens, Dermatophagoides / Q000737:chemistry* / Q000235:genetics; D060829:Arthropod Proteins; D003546:Cysteine Endopeptidases / Q000737:chemistry* / Q000235:genetics; D040002:Dermatophagoides pteronyssinus / Q000201:enzymology* / Q000235:genetics; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000737:chemistry* / Q000235:genetics; D006031:Glycosylation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012697:Serine Endopeptidases
D039741:Antigens, Dermatophagoides; D060829:Arthropod Proteins; D004792:Enzyme Precursors; D011994:Recombinant Proteins; C451165:Dermatophagoides pteronyssinus antigen p 3; D012697:Serine Endopeptidases; D003546:Cysteine Endopeptidases; C451162:Dermatophagoides pteronyssinus antigen p 1