Dehydrin Client Proteins Identified Using Phage Display Affinity Selected Libraries Processed With Paired-End Phage Sequencing.

Autor:	Unêda-Trevisoli SH; Department of Horticulture, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA; Department of Crop Production, São Paulo State University (Unesp), School of Agricultural and Veterinarian Sciences, São Paulo, Brazil., Dirk LMA; Department of Horticulture, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA., Carlos Bezerra Pereira FE; Department of Horticulture, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA; Department of Crop Production, São Paulo State University (Unesp), School of Agricultural and Veterinarian Sciences, São Paulo, Brazil; Pastotech Pasture Seeds, Campo Grande, Mato Grosso do Sul, Brazil., Chakrabarti M; School of Integrative Biological and Chemical Sciences, University of Texas Rio Grande Valley, Edinburg, Texas, USA., Hao G; Department of Plant and Soil Science, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Catalent Pharma Solution, Baltimore, Maryland, USA., Campbell JM; Department of Horticulture, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA; University of Kentucky Agricultural and Medical Biotechnology Program, Lexington, Kentucky, USA; Department of Toxicology and Cancer Biology, College of Medicine, University of Kentucky, Lexington, Kentucky, USA., Bassetti Nayakwadi SD; Department of Horticulture, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA; University of Kentucky Agricultural and Medical Biotechnology Program, Lexington, Kentucky, USA., Morrison A; Department of Horticulture, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA; University of Kentucky Agricultural and Medical Biotechnology Program, Lexington, Kentucky, USA., Joshi S; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA; Department of Plant and Soil Science, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Kentucky Tobacco Research and Development Center, Lexington, Kentucky, USA., Perry SE; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA; Department of Plant and Soil Science, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA., Sharma V; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA; Department of Plant and Soil Science, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA., Mensah C; Department of Horticulture, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA; Carter G. Woodson Academy, Fayette County Public Schools (FCPS), Lexington, Kentucky, USA., Willard B; Department of Horticulture, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA., de Lorenzo L; Department of Plant and Soil Science, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Department of Biochemistry and Molecular Biology, University of New Mexico, School of Medicine, Albuquerque, New Mexico, USA., Afroza B; Department of Horticulture, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA; Division of Vegetable Science, SKUAST- Kashmir, Srinagar, Kashmir, India., Hunt AG; Department of Plant and Soil Science, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA., Kawashima T; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA; Department of Plant and Soil Science, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA., Vaillancourt L; Department of Plant Pathology, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA., Pinheiro DG; Department of Crop Production, São Paulo State University (Unesp), School of Agricultural and Veterinarian Sciences, São Paulo, Brazil; Department of Agricultural, Livestock and Environmental Biotechnology, São Paulo State University (UNESP), School of Agricultural and Veterinary Sciences, Jaboticabal, São Paulo, Brazil., Downie AB; Department of Horticulture, Martin-Gatton College of Agriculture, Food and Environment, University of Kentucky, Lexington, Kentucky, USA; Seed Biology Program, University of Kentucky, Lexington, Kentucky, USA. Electronic address: adownie@uky.edu.
Jazyk:	angličtina
Zdroj:	Molecular & cellular proteomics : MCP [Mol Cell Proteomics] 2024 Dec; Vol. 23 (12), pp. 100867. Date of Electronic Publication: 2024 Oct 21.
DOI:	10.1016/j.mcpro.2024.100867
Abstrakt:	The late embryogenesis abundant proteins (LEAPs) are a class of noncatalytic, intrinsically disordered proteins with a malleable structure. Some LEAPs exhibit a protein and/or membrane binding capacity and LEAP binding to various targets has been positively correlated with abiotic stress tolerance. Regarding the LEAPs' presumptive role in protein protection, identifying client proteins (CtPs) to which LEAPs bind is one practicable means of revealing the mechanism by which they exert their function. To this end, we used phage display affinity selection to screen libraries derived from Arabidopsis thaliana seed mRNA with recombinant orthologous LEAPs from Arabidopsis and soybean (Glycine max). Subsequent high-throughput sequencing of DNA from affinity-purified phage was performed to characterize the entire subpopulation of phage retained by each LEAP ortholog. This entailed cataloging in-frame fusions, elimination of false positives, and aligning the hits on the CtP scaffold to reveal domains of respective CtPs that bound to orthologous LEAPs. This approach (paired-end phage sequencing) revealed a subpopulation of the proteome constituting the CtP repertoire in common between the two dehydrin orthologs (LEA14 and GmPm12) compared to bovine serum albumin (unrelated binding control). The veracity of LEAP:CtP binding for one of the CtPs (LEA14 and GmPM12 self-association) was independently assessed using temperature-related intensity change analysis. Moreover, LEAP:CtP interactions for four other CtPs were confirmed in planta using bimolecular fluorescence complementation assays. The results provide insights into the involvement of the dehydrin Y-segments and K-domains in protein binding. Competing Interests: Conflict of interest The authors declare no competing interests. (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu