The Role of SurA PPIase Domains in Preventing Aggregation of the Outer-Membrane Proteins tOmpA and OmpT. Issue 6 (15th March 2019)

• Record Type: Journal Article
• Authors: Humes, Julia R.; Schiffrin, Bob; Calabrese, Antonio N.; Higgins, Anna J.; Westhead, David R.; Brockwell, David J.; Radford, Sheena E.
• Is Part Of: Journal of molecular biology. Volume 431:Issue 6(2019)
• Page Start: 1267
• Page End: 1283
• Publication Date: 2019-03-15
• Subjects: OMP outer-membrane protein -- PPIase peptidylprolyl isomerase -- SurA ΔP2 SurA in which PPIase domain P2 has been deleted -- SurA N-Ct SurA in which both PPIase domains P1 and P2 are deleted -- BAM β-barrel assembly machinery; outer-membrane protein -- chaperone -- aggregation -- prolyl isomerase -- SurA; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2019.01.032
• Languages: English
• ISSNs: 0022-2836

Abstract

Abstract: SurA is a conserved ATP-independent periplasmic chaperone involved in the biogenesis of outer-membrane proteins (OMPs). Escherichia coli SurA has a core domain and two peptidylprolyl isomerase (PPIase) domains, the role(s) of which remain unresolved. Here we show that while SurA homologues in early proteobacteria typically contain one or no PPIase domains, the presence of two PPIase domains is common in SurA in later proteobacteria, implying an evolutionary advantage for this domain architecture. Bioinformatics analysis of > 350, 000 OMP sequences showed that their length, hydrophobicity and aggregation propensity are similar across the proteobacterial classes, ruling out a simple correlation between SurA domain architecture and these properties of OMP sequences. To investigate the role of the PPIase domains in SurA activity, we deleted one or both PPIase domains from E. coli SurA and investigated the ability of the resulting proteins to bind and prevent the aggregation of tOmpA (19 kDa) and OmpT (33 kDa). The results show that wild-type SurA inhibits the aggregation of both OMPs, as do the cytoplasmic OMP chaperones trigger factor and SecB. However, while the ability of SurA to bind and prevent tOmpA aggregation does not depend on its PPIase domains, deletion of even a single PPIase domain ablates the ability of SurA to prevent OmpT aggregation. The results demonstrate that the core domain of SurA endows its generic chaperone ability, while the presence of PPIase domains enhances its chaperone activity for specific OMPs, suggesting one reason for the conservation of multiple PPIase domains in SurA in proteobacteria. Graphical Abstract: Unlabelled Image Highlights: The role(s) of the two PPIase domains in E. coli SurA remains unresolved. Multiple SurA PPIase domains are conserved in β- and γ-proteobacteria. OMP sequence properties do not reveal a correlation with SurA domain architecture. tOmpA and OmpT require different SurA domains for chaperoning. SurA-OMP specificity suggests one reason for multiple PPIase domain conservation.