TY - JOUR
T1 - Purification of a glycoprotein elicitor of phytoalexin formation from Verticillium dahliae
AU - Davis, D. A.
AU - Low, P. S.
AU - Heinstein, P.
N1 - Funding Information:
* Supported by NSF Grant DCB 9005173. ‹Present Address: NHLBI; National Institutes of Health; Bethesda, MD 20892, U.S.A. ŒAddress all correspondence to: Peter Heinstein, Department of Medicinal Chemistry and Mol. Pharmacology, Purdue University, West Lafayette, IN. 47907-1333, U.S.A. Abbreviations used in text: AP-elicitor, acetone precipitated crude elicitor; BSA, bovine serum albumin; Con-A, concanavalin A; PNGase F, peptide N-glycosidase F; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.
Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 1998/4
Y1 - 1998/4
N2 - A glycoprotein, isolated from culture fluids of Verticillium dahliae was found to be a heat stable elicitor of phytoalexin biosynthesis in cotton cell suspension cultures. The elicitor was purified utilizing anion exchange, size exclusion, and Concanavalin-A (Con-A) chromatography in addition to electroelution from SDS-PAGE. The 65 kDa protein could be deglycosylated to a protein of molecular mass 53 kDa as determined by SDS-PAGE. Elicitor activity from the culture fluids as well as the Con-A chromatography purified elicitor was completely abolished by protease treatment, but not by periodate treatment or peptide N-glycosidase F (PNGase-F) digestion, indicating that only protein components are responsible for the elicitation of phytoalexins in cotton cells. The purified 65 kDa protein did not elicit H2O2 formation in contrast to the crude elicitor, which elicited both phytoalexin accumulation and the oxidative burst. The 65-kDa species was found to aggregate to a high molecular weight protein which retained some of its phytoalexin-inducing activity. The glycoprotein elicitor should prove useful in identifying second messengers leading to induction of phytoalexin biosynthesis.
AB - A glycoprotein, isolated from culture fluids of Verticillium dahliae was found to be a heat stable elicitor of phytoalexin biosynthesis in cotton cell suspension cultures. The elicitor was purified utilizing anion exchange, size exclusion, and Concanavalin-A (Con-A) chromatography in addition to electroelution from SDS-PAGE. The 65 kDa protein could be deglycosylated to a protein of molecular mass 53 kDa as determined by SDS-PAGE. Elicitor activity from the culture fluids as well as the Con-A chromatography purified elicitor was completely abolished by protease treatment, but not by periodate treatment or peptide N-glycosidase F (PNGase-F) digestion, indicating that only protein components are responsible for the elicitation of phytoalexins in cotton cells. The purified 65 kDa protein did not elicit H2O2 formation in contrast to the crude elicitor, which elicited both phytoalexin accumulation and the oxidative burst. The 65-kDa species was found to aggregate to a high molecular weight protein which retained some of its phytoalexin-inducing activity. The glycoprotein elicitor should prove useful in identifying second messengers leading to induction of phytoalexin biosynthesis.
UR - http://www.scopus.com/inward/record.url?scp=0032053233&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032053233&partnerID=8YFLogxK
U2 - 10.1006/pmpp.1998.0150
DO - 10.1006/pmpp.1998.0150
M3 - Article
AN - SCOPUS:0032053233
VL - 52
SP - 259
EP - 273
JO - Physiological and Molecular Plant Pathology
JF - Physiological and Molecular Plant Pathology
SN - 0885-5765
IS - 4
ER -