Structural characterization and mechanical properties of chimeric Masp1/Flag minispidroins

Xu, Shouying; Li, Xue; Zhou, Yizhong; Lin, Ying; Meng, Qing

doi:10.1016/j.biochi.2019.11.014

Cited by 17 publications

(16 citation statements)

References 49 publications

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“…Meanwhile, the molecular weight of Flag R -AcSp1 R is smaller than those of NTR 4 CT (42. (51.5 MPa [14,15] ), supporting the perspective that the recombinant spidroins with small molecular weight can also show excellent properties in terms of strength. [15,25] Therefore, reasonable design and combination of repetitive modules of recombinant spidroins could have better performance than the uniform iterated repetitive modules.…”

Section: Mechanical Properties Of the Chimeric Fiberssupporting

confidence: 59%

“…[29,30] Therefore, the excellent strength of Flag R-AcSp1 R can be explained by the higher content of β-sheet structure. [12,14,15] 4 | CONCLUSIONS F I G U R E 6 Secondary structures of silk-like fibers determined by FTIR spectroscopy. Decomposition of chimeric spider silk fiber spectrum in the amide I region (1600 1700 cm −1 ) Red peaks correspond to α-helix; blue peaks correspond to β-sheet; green peaks correspond to β-turn; pink peaks correspond to random coil; black curve corresponds to the original curve and orange curve corresponds to the sum of the decomposed spectra…”

Section: Structural Changes In Fiber Formationmentioning

confidence: 99%

“…[ 12–14 ] However, the related reports of spidroins mainly focused on the spidroins fused with Flag and MaSp, or the combination of some short motifs of different types of spidroins. [ 15–17 ] A previous study has shown that the fibers spun from the chimeric spidroin composed of AcSp1 repetitive modules and dragline repeat units exhibited better mechanical properties than fibers from spidroins only consisting of AcSp1 repetitive modules. With the increasing amounts of MaSp repeat unit, the mechanical properties of fibers decreased.…”

Section: Introductionmentioning

confidence: 99%

“…And the latest research showed that the repetitive units in chimeric spider silk sequences conferred strength and elasticity of fibers. [ 15 ] Here, we constructed a recombinant protein (Flag R ‐AcSp1 R ) comprised of a similar‐sized repetitive module from Flag and a repetitive module from AcSp1. After optimizing the expression and purification conditions, we got the chimeric protein without any extra tag.…”

Section: Introductionmentioning

confidence: 99%

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Expression and characterization of chimeric spidroins from flagelliform‐aciniform repetitive modules

2020

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Spiders can produce up to seven different types of silks or glues with different mechanical properties. Of these, flagelliform (Flag) silk is the most elastic, and aciniform (AcSp1) silk is the toughest. To produce a chimeric spider silk (spidroin) Flag R-AcSp1 R , we fused one repetitive module of flagelliform silk from Araneus ventricosus and one repetitive module of aciniform silk from Argiope trifasciata. The recombinant protein expressed in E. coli formed silk-like fibers by manual-drawing. CD analysis showed that the secondary structure of Flag R-AcSp1 R spidroin remained stable during the gradual reduction of pH from 7.0 to 5.5. The spectrum of FTIR indicated that the secondary structure of Flag R-AcSp1 R changed from α-helix to β-sheet. The conformation change of Flag R-AcSp1 R was similar to other spidroins in the fiber formation process. SEM analysis revealed that the mean diameter of the fibers was around 1 2 μm, and the surface was smooth and uniform. The chimeric fibers exhibited superior toughness (33.1 MJ/m 3) and tensile strength (261.4 MPa). This study provides new insight into design of chimeric spider silks with high mechanical properties.

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Section: Mechanical Properties Of the Chimeric Fiberssupporting

confidence: 59%

Section: Structural Changes In Fiber Formationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Expression and characterization of chimeric spidroins from flagelliform‐aciniform repetitive modules

2020

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“…By manipulating both the ratio of different motifs and combining different repetitive motifs derived from various spidroins, it is possible to design novel synthetic model proteins to understand and probe a broader range of artificial silk properties, including new specific biological materials with tunable mechanical properties. Examples of chimeric spider silk proteins include MaSp1/MaSp2 fusion proteins [44][45][46], MaSp1/Flag [47], MaSp2/Flag [48,49] or MaSp/AcSp [50]. Another example is the generation of chimeric silkworm-spider silks through expression in transgenic silkworms.…”

Section: Introductionmentioning

confidence: 99%

Expanding Canonical Spider Silk Properties through a DNA Combinatorial Approach

et al. 2020

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The properties of native spider silk vary within and across species due to the presence of different genes containing conserved repetitive core domains encoding a variety of silk proteins. Previous studies seeking to understand the function and material properties of these domains focused primarily on the analysis of dragline silk proteins, MaSp1 and MaSp2. Our work seeks to broaden the mechanical properties of silk-based biomaterials by establishing two libraries containing genes from the repetitive core region of the native Latrodectus hesperus silk genome (Library A: genes masp1, masp2, tusp1, acsp1; Library B: genes acsp1, pysp1, misp1, flag). The expressed and purified proteins were analyzed through Fourier Transform Infrared Spectrometry (FTIR). Some of these new proteins revealed a higher portion of β-sheet content in recombinant proteins produced from gene constructs containing a combination of masp1/masp2 and acsp1/tusp1 genes than recombinant proteins which consisted solely of dragline silk genes (Library A). A higher portion of β-turn and random coil content was identified in recombinant proteins from pysp1 and flag genes (Library B). Mechanical characterization of selected proteins purified from Library A and Library B formed into films was assessed by Atomic Force Microscopy (AFM) and suggested Library A recombinant proteins had higher elastic moduli when compared to Library B recombinant proteins. Both libraries had higher elastic moduli when compared to native spider silk proteins. The preliminary approach demonstrated here suggests that repetitive core regions of the aforementioned genes can be used as building blocks for new silk-based biomaterials with varying mechanical properties.

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Strategies for Making High‐Performance Artificial Spider Silk Fibers

Schmuck,

Greco,

Pessatti

et al. 2023

Adv Funct Materials

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Artificial spider silk is an attractive material for many technical applications since it is a biobased fiber that can be produced under ambient conditions but still outcompetes synthetic fibers (e.g., Kevlar) in terms of toughness. Industrial use of this material requires bulk‐scale production of recombinant spider silk proteins in heterologous host and replication of the pristine fiber's mechanical properties. High molecular weight spider silk proteins can be spun into fibers with impressive mechanical properties, but the production levels are too low to allow commercialization of the material. Small spider silk proteins, on the other hand, can be produced at yields that are compatible with industrial use, but the mechanical properties of such fibers need to be improved. Here, the literature on wet‐spinning of artificial spider silk fibers is summarized and analyzed with a focus on mechanical performance. Furthermore, several strategies for how to improve the properties of such fibers, including optimized protein composition, smarter spinning setups, innovative protein engineering, chemical and physical crosslinking as well as the incorporation of nanomaterials in composite fibers, are outlined and discussed.

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Structural characterization and mechanical properties of chimeric Masp1/Flag minispidroins

Cited by 17 publications

References 49 publications

Expression and characterization of chimeric spidroins from flagelliform‐aciniform repetitive modules

Expression and characterization of chimeric spidroins from flagelliform‐aciniform repetitive modules

Expanding Canonical Spider Silk Properties through a DNA Combinatorial Approach

Strategies for Making High‐Performance Artificial Spider Silk Fibers

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