Chimeric Spider Silk Proteins Mediated by Intein Result in Artificial Hybrid Silks

被引:13
作者
Lin, Senzhu [1 ,2 ]
Chen, Gefei [1 ,3 ]
Liu, Xiangqin [2 ]
Meng, Qing [1 ]
机构
[1] Donghua Univ, Inst Biol Sci & Biotechnol, Shanghai 201620, Peoples R China
[2] Dalhousie Univ, Dept Biochem & Mol Biol, Halifax, NS B3H 4R2, Canada
[3] Karolinska Inst, Ctr Alzheimer Res, Dept Neurobiol Care Sci & Soc NVS, S-14157 Stockholm, Sweden
基金
加拿大自然科学与工程研究理事会;
关键词
spider silk; intein; chimeric silk protein; hybrid silk; C-TERMINAL DOMAIN; WRAPPING SILK; MECHANICAL-PROPERTIES; REPETITIVE DOMAIN; SPIDROIN; DRAGLINE; FIBERS; SEQUENCE; GENE; ARANEAE;
D O I
10.1002/bip.22828
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Hybrid silks hold a great potential as specific biomaterials due to its controlled mechanical properties. To produce fibers with tunable properties, here we firstly made chimeric proteins in vitro, called W2C4CTand W2C8CT, with ligation of MaSp repetitive modules (C) with AcSp modules (W) by intein trans splicing technology from smaller precursors without final yield reduction. Intein mediated chimeric proteins form fibers at a low concentration of 0.4 mg/mL in 50 mM K3PO4 pH 7.5 just drawn by hand. Hybrid fibers show smoother surface, and also have stronger chemical resistance as compared with fibers from W2CT (W fibers) and mixture of W2CT/C8CT (MHF8 fibers). Fibers from chimeric protein W2C4CT (HFH4) have improved mechanical properties than W fibers; however, with more C modules W2C8CT fibers (HFH8) properties decreased, indicates the length proportion of various modules is very important and should be optimized for fibers with specific properties. Generally, hybrid silks generated via chimeric proteins, which can be simplified by intein trans splicing, has greater potential to produce fibers with tunable properties. Our research shows that intein mediated directional protein ligation is a novel way to make large chimeric spider silk proteins and hybrid silks. (C) 2016 Wiley Periodicals, Inc.
引用
收藏
页码:385 / 392
页数:8
相关论文
共 42 条
[1]   Carbonic Anhydrase Generates CO2 and H+ That Drive Spider Silk Formation Via Opposite Effects on the Terminal Domains [J].
Andersson, Marlene ;
Chen, Gefei ;
Otikovs, Martins ;
Landreh, Michael ;
Nordling, Kerstin ;
Kronqvist, Nina ;
Westermark, Per ;
Jornvall, Hans ;
Knight, Stefan ;
Ridderstrale, Yvonne ;
Holm, Lena ;
Meng, Qing ;
Jaudzems, Kristaps ;
Chesler, Mitchell ;
Johansson, Jan ;
Rising, Anna .
PLOS BIOLOGY, 2014, 12 (08)
[2]   Structural Model for the Spider Silk Protein Spidroin-1 [J].
Aparecido dos Santos-Pinto, Jose Roberto ;
Arcuri, Helen Andrade ;
Priewalder, Helga ;
Salles, Heliana Clara ;
Palma, Mario Sergio ;
Lubec, Gert .
JOURNAL OF PROTEOME RESEARCH, 2015, 14 (09) :3859-3870
[3]   Blueprint for a High-Performance Biomaterial: Full-Length Spider Dragline Silk Genes [J].
Ayoub, Nadia A. ;
Garb, Jessica E. ;
Tinghitella, Robin M. ;
Collin, Matthew A. ;
Hayashi, Cheryl Y. .
PLOS ONE, 2007, 2 (06)
[4]   Ancient Properties of Spider Silks Revealed by the Complete Gene Sequence of the Prey-Wrapping Silk Protein (AcSp1) [J].
Ayoub, Nadia A. ;
Garb, Jessica E. ;
Kuelbs, Amanda ;
Hayashi, Cheryl Y. .
MOLECULAR BIOLOGY AND EVOLUTION, 2013, 30 (03) :589-601
[5]   Fibre science - Supercontraction stress in wet spider dragline [J].
Bell, FI ;
McEwen, IJ ;
Viney, C .
NATURE, 2002, 416 (6876) :37-37
[6]   Full-Length Minor Ampullate Spidroin Gene Sequence [J].
Chen, Gefei ;
Liu, Xiangqin ;
Zhang, Yunlong ;
Lin, Senzhu ;
Yang, Zijiang ;
Johansson, Jan ;
Rising, Anna ;
Meng, Qing .
PLOS ONE, 2012, 7 (12)
[7]  
Foelix R.F., 1996, BIOL OF SPIDER
[8]   CANNIBALISM IN NATURAL-POPULATIONS [J].
FOX, LR .
ANNUAL REVIEW OF ECOLOGY AND SYSTEMATICS, 1975, 6 :87-106
[9]   Conserved C-Terminal Domain of Spider Tubuliform Spidroin 1 Contributes to Extensibility in Synthetic Fibers [J].
Gnesa, Eric ;
Hsia, Yang ;
Yarger, Jeffery L. ;
Weber, Warner ;
Lin-Cereghino, Joan ;
Lin-Cereghino, Geoff ;
Tang, Simon ;
Agari, Kimiko ;
Vierrat, Craig .
BIOMACROMOLECULES, 2012, 13 (02) :304-312
[10]   A conserved spider silk domain acts as a molecular switch that controls fibre assembly [J].
Hagn, Franz ;
Eisoldt, Lukas ;
Hardy, John G. ;
Vendrely, Charlotte ;
Coles, Murray ;
Scheibel, Thomas ;
Kessler, Horst .
NATURE, 2010, 465 (7295) :239-U131