title: Conformational and functional studies of a cytosolic 90 kDa heat shock protein Hsp90 from sugarcane.
abstract: Hsp90s are involved in several cellular processes, such as signaling, proteostasis, epigenetics, differentiation and stress defense. Although Hsp90s from different organisms are highly similar, they usually have small variations in conformation and function. Thus, the characterization of different Hsp90s is important to gain insight into the structure-function relationship that makes these chaperones key regulators in protein homeostasis. This work describes the characterization of a cytosolic Hsp90 from sugarcane and its comparison with Hsp90s from other plants. Previous expressed sequence tag (EST) studies in Saccharum spp. (sugarcane) predicted the presence of an mRNA coding for a cytosolic Hsp90. The corresponding cDNA was cloned, and the recombinant protein was purified and its conformation and function characterized. The structural conformation of Hsp90 was assessed by chemical cross-linking and hydrogen/deuterium exchange using mass spectrometry and hydrodynamic assays, which revealed regions accessible to solvent and that Hsp90 is an elongated dimer in solution. The in vivo expression of Hsp90 in sugarcane leaves was confirmed by western blot, and in vitro functional characterization indicated that sugarcane Hsp90 has strong chaperone activity.
authors.affiliation: Institute of Chemistry, University of Campinas-UNICAMP, P.O. Box 6154, 13083-970 Campinas, SP, Brazil.
authors.forename: Viviane C H
authors.initials: VC
authors.lastname: da Silva
authors.forename: Thiago C
authors.initials: TC
authors.lastname: Cagliari
authors.forename: Tatiani B
authors.initials: TB
authors.lastname: Lima
authors.forename: Fábio C
authors.initials: FC
authors.lastname: Gozzo
authors.forename: Carlos H I
authors.initials: CH
authors.lastname: Ramos
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003600:Cytosol / Q000378:metabolism*; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D031786:Saccharum / Q000737:chemistry*; D017386:Sequence Homology, Amino Acid
chemical_list: D018841:HSP90 Heat-Shock Proteins; D018832:Molecular Chaperones; D010940:Plant Proteins