title: Structure-activity studies of GmSubPep, a soybean peptide defense signal derived from an extracellular protease.
abstract: GmSubPep, a 12-amino acid peptide isolated from soybean leaves, induces the expression of genes in soybean suspension-cultured cells that encode proteins involved in defense against pathogens. The peptide is derived from an extracellular subtilisin-like protease (subtilase) and binds a putative cell-surface receptor that initiates a defense signaling cascade. Interaction of the peptide with its receptor results in alkalinization of soybean suspension cell media which can be utilized to analyze the kinetics of receptor binding. Substitutions of alanine at each of the 12 amino acid positions revealed that the amino acids at positions 10 (arginine) and 12 (histidine) were essential for activity. Both analogs were able to reduce the physiological effects of GmSubPep associated with receptor binding. Deletion of the C-terminal histidine [GmSubPep(1-11)] abolished the alkalinizing activity and this peptide was also a strong competitor for receptor binding. Deletion of N-terminal amino acids from GmSubPep caused a sequential loss of activity with no alkalinizing activity for GmSubPep(4-12). However, the N-terminal deleted peptides did not compete with GmSubPep for receptor binding. Further modifications at the arginine-10 position indicated that an ionizable proton was not essential for activity as an attenuated response was found for a citrulline substitution. Substitution of the histidine-12 with methylated histidine at position N-1 of the imidazole group abolished activity, whereas substitution at N-3 was completely active, indicating that the N-3 analog retains important receptor binding properties. This study indicates that the extreme C-terminal of GmSubPep has important signal transduction properties while the C-terminal is essential for receptor interaction.
authors.affiliation: Institute of Biological Chemistry, Washington State University, Pullman, WA 99164-6340, United States. pearce@wsu.edu
authors.forename: Gregory
authors.initials: G
authors.lastname: Pearce
authors.forename: Gerhard
authors.initials: G
authors.lastname: Munske
authors.forename: Yube
authors.initials: Y
authors.lastname: Yamaguchi
authors.forename: Clarence A
authors.initials: CA
authors.lastname: Ryan
mesh_terms: D002478:Cells, Cultured; D006639:Histidine / Q000737:chemistry; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018515:Plant Leaves / Q000201:enzymology / Q000378:metabolism; D011485:Protein Binding; D011956:Receptors, Cell Surface; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology; D030262:Soybean Proteins / Q000737:chemistry* / Q000378:metabolism*; D013025:Glycine max / Q000201:enzymology / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013381:Subtilisins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D010455:Peptides; D011956:Receptors, Cell Surface; D030262:Soybean Proteins; D006639:Histidine; D013381:Subtilisins; C119297:subtilase, plant