alexa The Binding of Ca2+-Dependent Protein Kinase to the Suppressor of Potato Late Blight Pathogen Proved by Fluorescence Correlation Spectroscopy (FCS) Inhibits the NADPH Oxidase and Active Oxygen Generation in Potato Cell
ISSN: 2157-7471

Journal of Plant Pathology & Microbiology
Open Access

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Research Article

The Binding of Ca2+-Dependent Protein Kinase to the Suppressor of Potato Late Blight Pathogen Proved by Fluorescence Correlation Spectroscopy (FCS) Inhibits the NADPH Oxidase and Active Oxygen Generation in Potato Cell

Naotaka Furuichi1*, Kazutoshi Yokokawa2, Koji Ohnishi1 and Masatoshi Ohta3

1Center for Transdisciplinary Research, Niigata University, Niigata 950-2181, Japan

2Graduate School of Science and Technology, Niigata University, Niigata 950-2181, Japan

3Super Oxide Radical Institute, Niigata Nishi-ku, Niigata 950-2102, Japan

*Corresponding Author:
Naotaka Furuichi
Center for Transdisciplinary Research
Niigata University
Niigata 950-2181, Japan
Tel: 81252627520
Fax: 81252627520
E-mail: [email protected]

Received date: July 04, 2015; Accepted date: August 05, 2015; Published date: August 10, 2015

Citation: Furuichi N, Yokokawa K, Ohnishi K, Ohta M (2015) The Binding of Ca2+- Dependent Protein Kinase to the Suppressor of Potato Late Blight Pathogen Proved by Fluorescence Correlation Spectroscopy (FCS) Inhibits the NADPH Oxidase and Active Oxygen Generation in Potato Cell. J Plant Pathol Microbiol S4:001. doi:10.4172/2157-7471.S4-001

Copyright: © 2015 Furuichi N, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

 

Abstract

Representing the suppressor for hypersensitive cell death of plant cell, glucan from Phytophthora infestans (Pi) was reported, and that the suppressor inhibited the accumulation of phytoalexin and hypersensitive cell death. To evaluate the activation of calcium dependent protein kinase (CDPK) after the binding of suppressor from Pi, fluorescence correlation spectroscopy (FCS) was applied to single GFP-CDPK and Alexa labeled suppressor. We constructed chimeric protein, tandemly fused green FP (GFP)-CDPK and suppressor with the alexa 633 labeled-suppressor antibodies (Abs). Dual color FCS provides information about the coincidence of spectrally two fluorescent molecules at a single-molecule level. Here we report that for the inhibition of NADPH oxidase of potato, the suppressor bound to CDPK peptide (kinase domain-I and -III peptides), CDPK phosphorylated the NADPH oxidase in gel kinase assay, and that the generation of active oxygen by NADPH oxydase was inhibited by the suppressor. This means HR inhibiting suppressor from the plant pathogen control the superoxide radical formation in plant cell by signal of the phosphorylation by a CDPK.

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