Structural Basis for pH-mediated Regulation of F-actin Severing by Gelsolin Domain 1

Fan, Jing; Goh, Honzhen; Ding, Ke; Xue, Bin; Robinson, Robert; Yang, Daiwen

doi:10.1038/srep45230

Cited by 8 publications

(7 citation statements)

References 38 publications

(52 reference statements)

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“…A similar observation was reported in a recent multi-dimensional NMR experiments based study where 28–161 was studied at two pH values, 7.3 and 5 and no nuclear Overhauser effects (NOEs) could be recorded for the residues 145–161 under either condition. This was attributed to the highly disordered nature of these residues, and thus the orientation of the tail could not be defined 19 . In our crystal structure, four Ca 2+ ions were present in the entire assembly, two bound to each monomer, including both Type I (G 65 , E 97 , V 145 ) and Type II (D 109 , G 114 , A 116 ) Ca 2+ binding sites.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Bonsai Gelsolin Survives Heat Induced Denaturation by Forming β-Amyloids which Leach Out Functional Monomer

Badmalia

Sharma

Yadav

et al. 2018

Sci Rep

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Here, we report that minimal functional gelsolin i.e. fragment 28–161 can display F-actin depolymerizing property even after heating the protein to 80 °C. Small angle X-ray scattering (SAXS) data analysis confirmed that under Ca2+-free conditions, 28–161 associates into monomer to dimer and tetramer, which later forms β-amyloids, but in presence of Ca2+, it forms dimers which proceed to non-characterizable aggregates. The dimeric association also explained the observed decrease in ellipticity in circular dichroism experiments with increase in temperature. Importantly, SAXS data based models correlated well with our crystal structure of dimeric state of 28–161. Characterization of higher order association by electron microscopy, Congo red and ThioflavinT staining assays further confirmed that only in absence of Ca2+ ions, heating transforms 28–161 into β-amyloids. Gel filtration and other experiments showed that β-amyloids keep leaching out the monomer, and the release rates could be enhanced by addition of L-Arg to the amyloids. F-actin depolymerization showed that addition of Ca2+ ions to released monomer initiated the depolymerization activity. Overall, we propose a way to compose a supramolecular assembly which releases functional protein in sustained manner which can be applied for varied potentially therapeutic interventions.

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Section: Resultsmentioning

confidence: 99%

“…Little shape-function work has been performed on this domain alone though it can depolymerize F-actin more efficiently than native protein i.e. gelsolin 6 , 19 . While exploring thermal stability of gelsolin we recently delineated that gelsolin can also be activated by physiological temperature 10 .…”

Section: Discussionmentioning

confidence: 99%

Bonsai Gelsolin Survives Heat Induced Denaturation by Forming β-Amyloids which Leach Out Functional Monomer

Badmalia

Sharma

Yadav

et al. 2018

Sci Rep

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“…Therefore, the salt-bridge interaction should not be a dominant factor for the pH-responsive behavior. It is well established that the regulation of the protein structure and stability by environmental pH is achieved through protonation or deprotonation of acidic (negatively charged) and basic (positively charged) residues. , In the range of pH 5–7, only D63, D69, and H76 may change their charge states. Despite that D63N is less stable than WT, D63N and WT CTD Fl have similar sensitivities to pH (Figure A,C), indicating that D63 is not important for the pH-responsive property.…”

Section: Discussionmentioning

confidence: 99%

C-Terminal Domains of Spider Silk Proteins Having Divergent Structures but Conserved Functional Roles

Fan

Shi

et al. 2022

Biomacromolecules

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Spider silk is self-assembled from silk proteins or spidroins. C-terminal domains (CTDs) of various types of spidroins are relatively conserved in amino acid sequences and are suggested to adopt similar structures and perform similar functional roles in spidroin storage and silk formation. Here, we solved the structure of the CTD from a capture-spiral silk protein (CTDFl) and characterized its stability and fibril formation in the presence and absence of a reducing agent at different pH values. CTDFl adopts a dimeric structure with 8 helices, but the CTDs of other types of spidroins exist in a domain-swapped dimeric structure with 10 helices. Despite the structural differences, CTDFl is pH-responsive in stability and fibril formation, similar to the CTDs from minor and major ampullate spidroins. Thus, the functional role of CTDs in silk fiber formation seems conserved. Comparing wild-type CTDFl and its mutants, we found that the pH-responsive behavior results from the protonation of H76, which is conserved from different spider species. In addition, the fibril formation rate of CTDFl correlates with its instability, suggesting that structural changes are involved in fibril formation.

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“…Steady-state 15 N–{ 1 H} heteronuclear NOE values of CSDex were measured on a 700 MHz spectrometer at 298 K according to the previously described method ( 44 ). In the measurements, the recycle delay was 8 s and the proton saturation time was 4 s. The 15 N–{ 1 H} NOE values were derived from ratios of the peak intensities with and without proton saturation.…”

Section: Methodsmentioning

confidence: 99%

Structural basis of DNA binding to human YB-1 cold shock domain regulated by phosphorylation

Zhang

Fan

et al. 2020

Nucleic Acids Research

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Human Y-box binding protein 1 (YB-1) is a multifunctional protein and overexpressed in many types of cancer. It specifically recognizes DNA/RNA through a cold shock domain (CSD) and regulates nucleic acid metabolism. The C-terminal extension of CSD and the phosphorylation of S102 are indispensable for YB-1 function. Until now, the roles of the C-terminal extension and phosphorylation in gene transcription and translation are still largely unknown. Here, we solved the structure of human YB-1 CSD with a C-terminal extension sequence (CSDex). The structure reveals that the extension interacts with several residues in the conventional CSD and adopts a rigid structure instead of being disordered. Either deletion of this extension or phosphorylation of S102 destabilizes the protein and results in partial unfolding. Structural characterization of CSDex in complex with a ssDNA heptamer shows that all the seven nucleotides are involved in DNA–protein interactions and the C-terminal extension provides a unique DNA binding site. Our DNA-binding study indicates that CSDex can recognize more DNA sequences than previously thought and the phosphorylation reduces its binding to ssDNA dramatically. Our results suggest that gene transcription and translation can be regulated by changing the affinity of CSDex binding to DNA and RNA through phosphorylation, respectively.

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Structural Basis for pH-mediated Regulation of F-actin Severing by Gelsolin Domain 1

Cited by 8 publications

References 38 publications

Bonsai Gelsolin Survives Heat Induced Denaturation by Forming β-Amyloids which Leach Out Functional Monomer

Bonsai Gelsolin Survives Heat Induced Denaturation by Forming β-Amyloids which Leach Out Functional Monomer

C-Terminal Domains of Spider Silk Proteins Having Divergent Structures but Conserved Functional Roles

Structural basis of DNA binding to human YB-1 cold shock domain regulated by phosphorylation

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