title: In vivo substrate diversity and preference of small heat shock protein IbpB as revealed by using a genetically incorporated photo-cross-linker.
abstract: Small heat shock proteins (sHSPs), as ubiquitous molecular chaperones found in all forms of life, are known to be able to protect cells against stresses and suppress the aggregation of a variety of model substrate proteins under in vitro conditions. Nevertheless, it is poorly understood what natural substrate proteins are protected by sHSPs in living cells. Here, by using a genetically incorporated photo-cross-linker (p-benzoyl-l-phenylalanine), we identified a total of 95 and 54 natural substrate proteins of IbpB (an sHSP from Escherichia coli) in living cells with and without heat shock, respectively. Functional profiling of these proteins (110 in total) suggests that IbpB, although binding to a wide range of cellular proteins, has a remarkable substrate preference for translation-related proteins (e.g. ribosomal proteins and amino-acyl tRNA synthetases) and moderate preference for metabolic enzymes. Furthermore, these two classes of proteins were found to be more prone to aggregation and/or inactivation in cells lacking IbpB under stress conditions (e.g. heat shock). Together, our in vivo data offer novel insights into the chaperone function of IbpB, or sHSPs in general, and suggest that the preferential protection on the protein synthesis machine and metabolic enzymes may dominantly contribute to the well known protective effect of sHSPs on cell survival against stresses.
authors.affiliation: From the State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871 and.
authors.forename: Xinmiao
authors.initials: X
authors.lastname: Fu
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Shi
authors.forename: Linxuan
authors.initials: L
authors.lastname: Yan
authors.forename: Hanlin
authors.initials: H
authors.lastname: Zhang
authors.forename: Zengyi
authors.initials: Z
authors.lastname: Chang
mesh_terms: D003432:Cross-Linking Reagents / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D018869:Heat-Shock Response / Q000502:physiology*; D014176:Protein Biosynthesis / Q000502:physiology*
chemical_list: D003432:Cross-Linking Reagents; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; C097579:IbpB protein, E coli
keywords: Heat Shock Protein; Metabolic Enzymes; Molecular Chaperone; Photocross-linking; Protein Aggregation; Protein Folding; Ribosomes; Small Heat Shock Proteins; Translation; Unnatural Amino Acid
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