Residues Phe103 and Phe149 are critical for the co-chaperone activity of Bacillus licheniformis GrpE

Lin, Min-Guan; Chi, Meng-Chun; Chen, Bo-En; Wang, Tzu-Fan; Lo, Huei-Fen; Lin, Long-Liu

doi:10.1016/j.ijbiomac.2014.09.013

Cited by 2 publications

(3 citation statements)

References 41 publications

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“…This mutation enhanced the thermal stability and functional activity of the protein and augmented the thermotolerance of yeast cells. Mutational studies on GrpE of Bacillus licheniformis show that phenylalanine residues at the four‐helical bundle and beta‐sheet wing control the side chain packing and regulate thermal stability 49 . Multiple sequence alignment of GrpE from mesophilic bacteria (Figure S7) shows that mesophiles lacking aromatic residues at F91/F137 positions share less than 35% sequence identity with E. coli GrpE 50,51 .…”

Section: Discussionmentioning

confidence: 99%

“…Mutational studies on GrpE of Bacillus licheniformis show that phenylalanine residues at the four-helical bundle and beta-sheet wing control the side chain packing and regulate thermal stability. 49 Multiple sequence alignment of GrpE from mesophilic bacteria (Figure S7) shows that mesophiles lacking aromatic residues at F91/F137 positions share less than 35% sequence identity with E. coli GrpE. 50,51 This is in comparison to >80% sequence identity in the ones that have the aromatic residues (Figures 1C and S7).…”

Section: Introduction Of Charged Residues In the Aromatic Core Does N...mentioning

confidence: 99%

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Investigating the functional role of a buried interchain aromatic cluster in Escherichia coli GrpE dimer

et al. 2022

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Aromatic clusters in the core of proteins are often involved in imparting structural stability to proteins. However, their functional importance is not always clear. In this study, we investigate the thermosensing role of a phenylalanine cluster present in the GrpE homodimer. GrpE, which acts as a nucleotide exchange factor for the molecular chaperone DnaK, is well known for its thermosensing activity resulting from temperature-dependent structural changes that allow control of chaperone function. Using mutational analysis, we show that an interchain phenylalanine cluster in a four-helix bundle of the GrpE homodimer assists in the thermosensing ability of the co-chaperone. Substitution of aromatic residues with hydrophobic ones in the core of the four-helix bundle reduces the thermal stability of the bundle and that of a connected coiled-coil domain, which impacts thermosensing. Cell growth assays and SEM images of the mutants show filamentous growth of Escherichia coli cells at 42 C, which corroborates with the defect in thermosensing. Our work suggests that the interchain edge-to-face aromatic cluster is important for the propagation of the structural signal from the coiled-coil domain to the four-helical bundle of GrpE, thus facilitating GrpE-mediated thermosensing in bacteria.

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Section: Discussionmentioning

confidence: 99%

Section: Introduction Of Charged Residues In the Aromatic Core Does N...mentioning

confidence: 99%

Investigating the functional role of a buried interchain aromatic cluster in Escherichia coli GrpE dimer

et al. 2022

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“…However, after Hsp70 consumes the ATP‐releasing peptide chain, GrpE depolymerizes into a monomer and then Hsp70 recombines ATP (David et al, 1998). Studies have shown that GrpE mutations in E. coli can affect heat shock protein function and regulate cell heat shock responses te cell heat shock responses (Lin et al, 2015). Mutation of the 167th histidine to leucine (H167L) in the GrpE protein enhances thermostability and high‐temperature resistance of E. coli (John & Delaney, 1990; Wu, Ang, Snavely, & Costa, 1994).…”

Section: Introductionmentioning

confidence: 99%

Identification of the nucleotide exchange factor BmGrpE and its role under high‐temperature stress in silkworm, Bombyx mori

Mao

et al. 2020

Arch Insect Biochem Physiol

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The high‐temperature stress gene GrpE plays an important role in coping with high‐temperature stress. The mutation of key sites of this gene can improve the high‐temperature resistance of organisms. In the present study, using complementary DNAs from the silkworm fat body as the template, the open reading frame sequence of the GrpE gene (BmGrpE) was amplified and was found to be 644 bp in length and encode a protein with a predicted molecular weight of 24.1 kDa. The presence of a binding site for the heat shock transcription factor (Hsf1) at −1440 bp upstream of its coding region indicates that BmGrpE may respond to high‐temperature stress. BmGrpE was constitutively expressed throughout developmental stages, with the highest level observed in the 5th instar larvae stage. Moreover, in 5th instar larvae (the 3th day), BmGrpE was expressed in all tissues examined, with the highest levels in the fat body, silk gland, and midgut. Interestingly, under high‐temperature stress, TiO2 nanoparticle treatment increased the messenger RNA levels of BmGrpE in the fat body and silk gland. After treatment with dsRNA of BmGrpE, the cell viability of BmN cells was significantly decreased under 34°C and H2O2 stress (p < .05). Mutation of BmGrpE (H163L) enhanced the resistance of BmN cells under high‐temperature stress. These results provide new clues for the study of molecular mechanisms of insect resistance to high temperatures.

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Residues Phe103 and Phe149 are critical for the co-chaperone activity of Bacillus licheniformis GrpE

Cited by 2 publications

References 41 publications

Investigating the functional role of a buried interchain aromatic cluster in Escherichia coli GrpE dimer

Investigating the functional role of a buried interchain aromatic cluster in Escherichia coli GrpE dimer

Identification of the nucleotide exchange factor BmGrpE and its role under high‐temperature stress in silkworm, Bombyx mori

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