A putative three-dimensional targeting motif of polygalacturonase (PehA), a protein secreted through the type II (GSP) pathway in Erwinia carotovora.
Intramolecular information specifying protein secretion through the type II (GSP) pathway of Gram-negative bacteria was investigated. Two regions of the polygalacturonase (PehA) of Erwinia carotovora containing residues proposed to be included in a targeting motif were located, one close to the C-terminus between residues 342 and 369 and another between residues 84 and 135 in the large central loops. The regions were required together to promote secretion. Further residues in the middle of the protein were required for proper positioning of the regions, suggesting that they were both involved in interaction with the GSP. To our knowledge, this is the first time that a possible three-dimensional targeting motif has been defined. At least one of the motifs comprises a cluster on the surface of the protein. The two motifs are structurally dissimilar, suggesting that there are two distinct recognition regions in the GSP apparatus. Finally, we propose that the targeting motifs are of a complex conformational nature with some variability accommodated, as illustrated by the observation that many mutations exhibited no clear phenotype individually but, in combination, severely compromised secretion.
Palomäki
Tiina
T
Department of Biosciences, Division of Genetics, Viikki Biocenter, University of Helsinki, PO Box 56, FIN-00014 Finland.
Pickersgill
Richard
R
Riekki
Ruusu
R
Romantschuk
Martin
M
Saarilahti
Hannu T
HT
D020816:Amino Acid Motifs; D008958:Models, Molecular; D009154:Mutation; D016973:Pectobacterium carotovorum / Q000737:chemistry / Q000378:metabolism*; D011096:Polygalacturonase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D011487:Protein Conformation
D011096:Polygalacturonase