Designing recombinant spider silk proteins to control assembly

Winkler, Stefan; Szela, Sandra; Avtges, Peter; Valluzzi, Regina; Kirschner, Daniel A.; Kaplan, David L.

doi:10.1016/s0141-8130(98)00088-9

Cited by 104 publications

(93 citation statements)

References 11 publications

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“…For example, tweaking the native or consensus sequences of spider silks to include encoded 'triggers' to regulate molecular-level assembly of the proteins has been demonstrated [17][18][19][20][21]. Chemical redox triggers, or a kinase recognition site for biochemical phosphorylation, flanking the b-sheet forming regions of the proteins have been demonstrated.…”

Section: Spider Silksmentioning

confidence: 99%

Silk-based biomaterials

et al. 2003

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Silk from the silkworm, Bombyx mori, has been used as biomedical suture material for centuries. The unique mechanical properties of these fibers provided important clinical repair options for many applications. During the past 20 years, some biocompatibility problems have been reported for silkworm silk; however, contamination from residual sericin (glue-like proteins) was the likely cause. More recent studies with well-defined silkworm silk fibers and films suggest that the core silk fibroin fibers exhibit comparable biocompatibility in vitro and in vivo with other commonly used biomaterials such as polylactic acid and collagen. Furthermore, the unique mechanical properties of the silk fibers, the diversity of side chain chemistries for 'decoration' with growth and adhesion factors, and the ability to genetically tailor the protein provide additional rationale for the exploration of this family of fibrous proteins for biomaterial applications. For example, in designing scaffolds for tissue engineering these properties are particularly relevant and recent results with bone and ligament formation in vitro support the potential role for this biomaterial in future applications. To date, studies with silks to address biomaterial and matrix scaffold needs have focused on silkworm silk. With the diversity of silk-like fibrous proteins from spiders and insects, a range of native or bioengineered variants can be expected for application to a diverse set of clinical needs. r

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Section: Spider Silksmentioning

confidence: 99%

Silk-based biomaterials

et al. 2003

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“…Winkler et al added methionine residues flanking the β-sheet forming polyalanine sequences of a recombinant silk protein. The oxidized methionine residues sterically hinder β-sheet formation and increase the protein's hydrophilicity, while in the reduced state, the added residues have no effect on crystallization [53]. They also modified a recombinant silk protein to include a phosphorylation site for cyclic AMP-dependent protein kinase (cAPK).…”

Section: Silk-like Polypeptides-mentioning

confidence: 99%

“…They also modified a recombinant silk protein to include a phosphorylation site for cyclic AMP-dependent protein kinase (cAPK). By introducing charged phosphate groups, the hydrophobic interactions required for β-sheet formation were interrupted, also preventing insolubility [53].…”

Section: Silk-like Polypeptides-mentioning

confidence: 99%

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Peptide-based biopolymers in biomedicine and biotechnology

Chow

Nunalee

Lim

et al. 2008

Materials Science and Engineering: R: Reports

286

231

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Peptides are emerging as a new class of biomaterials due to their unique chemical, physical, and biological properties. The development of peptide-based biomaterials is driven by the convergence of protein engineering and macromolecular self-assembly. This review covers the basic principles, applications, and prospects of peptide-based biomaterials. We focus on both chemically synthesized and genetically encoded peptides, including poly-amino acids, elastin-like polypeptides, silk-like polymers and other biopolymers based on repetitive peptide motifs. Applications of these engineered biomolecules in protein purification, controlled drug delivery, tissue engineering, and biosurface engineering are discussed.

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“…Proteins derived from genetic engineering are characterized by control over composition, sequence, molecular weight, and stereochemical purity. Motifs of silkworm and spider silk proteins have been studied in a similar manner to understand structures in different environments, and spider silk proteins have been modified for the purpose of controlled assembly (17)(18)(19)(20)(21)(22)(23)(24).…”

mentioning

confidence: 99%

Cloning, Expression, and Assembly of Sericin-like Protein

Huang

Valluzzi

Bini

et al. 2003

Journal of Biological Chemistry

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Recombinant sericin proteins of different molecular masses (17.4, 31.9, and 46.5 kDa), based on the 38-amino acid repetitive motif of native sericin, were cloned, expressed, and purified. The recombinant sericin self-assembled during dialysis (starting concentration of 2.5 mg/ml) forming twisted fibers. Circular dichroism and Fourier transform infrared spectroscopy studies demonstrated protein conformational transitions occurred from random coil to ␤-sheets during the dialysis. Congo red-stained recombinant sericin fibrils exhibited applegreen birefringence, indicating long-range order in the array of ␤-sheets. Biosynthetic sericin has a high content of polar amino acids (e.g. Silk of Bombyx mori is composed of two kinds of proteins: the fibroin which is synthesized in the posterior silk gland and the sericins produced by the middle silk gland. The sericins bind two fibroin fibers together in the anterior region of the gland as the fibers are spun to form the cocoon. Sericins represent a family of proteins whose molecular mass ranges from 20 to 310 kDa (2, 3) and are characterized by an unusually high serine content, 38.1 mol % (4). Sericins are soluble in hot water, which makes it possible to degum the silk fibroin threads.Two genes encode sericins, Ser1 and Ser2 (5-8). The different molecular weight sericins are the products of different splicing events at the transcript level. Some conformation studies with sericins from the silk gland of B. mori or regenerated sericin from B. mori cocoons suggested random coil with some ␤ structure (9, 10). However, the samples used in these studies were mixtures of the various native sericin proteins, therefore the structure of the individual proteins is unknown and the contribution of specific sequences toward secondary structure is not confirmed. Sericin model peptides such as poly(L-serine) (11-16), poly(O-benzyl-L-serine) (12), and poly(O-acetyl-L-serine) (13,14) were synthesized for structural feature characterization. The ability of poly(L-serine) to fold into a ␤ conformation depended on molecular weight. Low molecular weight poly(L-serine) (molecular weight 650) was soluble and adopted a random coil conformation in aqueous solution, whereas high molecular weight poly(L-serine) (molecular weight 105,000) formed ␤-sheets and was insoluble in water (11). However, homopolypeptides like poly(L-serine) do not match native sericin in terms of amino acid composition, sequence, and molecular weight. Therefore, to better understand sericin structure, interactions between sericin and fibroin, and the biological relevance of sericin in fiber structure, high molecular weight pure sericin-like proteins are required.Sericin 1 encoded by Ser 1 consists of 70 repeats of the 38-amino acid motif (5): SSTGSSSNTDSNSNSVGSSTS-GGSSTYGYSSNSRDGSV. The molecular mass of sericin 1 proteins is 76 -284 kDa. The 38-amino acid repeat motif exhibits a high content of hydrophilic amino acids: 44.7% serine, 10.5% threonine, and 7.9% tyrosine, very close to the average amino acid composition of serici...

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Designing recombinant spider silk proteins to control assembly

Cited by 104 publications

References 11 publications

Silk-based biomaterials

Silk-based biomaterials

Peptide-based biopolymers in biomedicine and biotechnology

Cloning, Expression, and Assembly of Sericin-like Protein

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