Synthesis and structural characterization of silk-like materials incorporated with an elastic motif

被引:20
作者
Yao, JM [1 ]
Asakura, T [1 ]
机构
[1] Tokyo Univ Agr & Technol, Dept Biotechnol, Tokyo 1848588, Japan
关键词
Bombyx mori silk fibroin; circular dichroism; genetic engineering synthesis; nuclear magnetic resonance; silk-like materials;
D O I
10.1093/jb/mvg014
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
Genetic engineering strategies were applied to synthesize silk-like materials, [(GVPGV)(2)GG(GAGAGS)(3)AS](n). The primary structure of these materials represents the repetitive crystalline region of Bombyx mori silk fibroins incorporated with an elastic motif selected from animal elastin. The oligonucleotides were designed to encode the desired recombinant proteins and then expressed in the Escherichi coli system. The expression and purification conditions for the production of the recombinant proteins were optimized. C-13 CP/MAS NMR was used for structural characterization in the solid state, where the isotope labeling was performed using a modified M9 medium. The secondary structures of these materials are primarily governed by the designated amino acid sequence, where the B. mori silk fibroin block, (GAGAGS)(3), tends to form the crystalline region, which is interrupted by the flexible (GVPGV)(2) block. The CD data suggested that the structure of these materials was length-dependent in the solution state, i.e., a higher molecule weight leads to a higher ordered structure.
引用
收藏
页码:147 / 154
页数:8
相关论文
共 38 条
[1]   Silk-based biomaterials [J].
Altman, GH ;
Diaz, F ;
Jakuba, C ;
Calabro, T ;
Horan, RL ;
Chen, JS ;
Lu, H ;
Richmond, J ;
Kaplan, DL .
BIOMATERIALS, 2003, 24 (03) :401-416
[2]  
AOTA S, 1988, NUCLEIC ACIDS RES, V16, P315
[3]   CONFORMATION CHARACTERIZATION OF BOMBYX-MORI SILK FIBROIN IN THE SOLID-STATE BY HIGH-FREQUENCY C-13 CROSS POLARIZATION MAGIC ANGLE SPINNING NMR, X-RAY-DIFFRACTION, AND INFRARED-SPECTROSCOPY [J].
ASAKURA, T ;
KUZUHARA, A ;
TABETA, R ;
SAITO, H .
MACROMOLECULES, 1985, 18 (10) :1841-1845
[4]   Heterogeneous structure of silk fibers from Bombyx mori resolved by 13C solid-state NMR spectroscopy [J].
Asakura, T ;
Yao, JM ;
Yamane, T ;
Umemura, K ;
Ulrich, AS .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2002, 124 (30) :8794-8795
[5]   Design, expression, and structural characterization of hybrid proteins of Samia cynthia ricini and Bombyx mori silk fibroins [J].
Asakura, T ;
Kato, H ;
Yao, J ;
Kishore, R ;
Shirai, M .
POLYMER JOURNAL, 2002, 34 (12) :936-943
[6]   13C CP/MAS NMR study on structural heterogeneity in Bombyx mori silk fiber and their generation by stretching [J].
Asakura, T ;
Yao, JM .
PROTEIN SCIENCE, 2002, 11 (11) :2706-2713
[7]   A repeated β-turn structure in poly(Ala-Gly) as a model for silk I of Bombyx mori silk fibroin studied with two-dimensional spin-diffusion NMR under off magic angle spinning and rotational echo double resonance [J].
Asakura, T ;
Ashida, J ;
Yamane, T ;
Kameda, T ;
Nakazawa, Y ;
Ohgo, K ;
Komatsu, K .
JOURNAL OF MOLECULAR BIOLOGY, 2001, 306 (02) :291-305
[8]  
Asakura T., 1994, Encyclopedia of AgriculturalScience, P1
[9]   Mechanism of stabilization of helical conformations of polypeptides by water containing trifluoroethanol [J].
CammersGoodwin, A ;
Allen, TJ ;
Oslick, SL ;
McClure, KF ;
Lee, JH ;
Kemp, DS .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1996, 118 (13) :3082-3090
[10]  
Cappello J., 1994, PLASTICS MICROBES, P35