Abstract
Bacteria were genetically engineered to produce two spider silk protein variants composed of basic repeat units combining a flagelliform elastic motif ([GPGGX]4) and a major ampullate silk strength motif ([linker/poly-alanine]. The secondary structures of the pure recombinant proteins in solution were determined by circular dichroism. The data presented suggest that the nature of the 5th and 10th amino acid (X) in the [GPGGX]2 elastic motif and temperature have an impact on the amount of β-sheet structures present in the proteins. More specifically, increasing temperatures seem to be positively correlated with β-sheet formation for both proteins and this state is irreversible or reversible when both X (5th and 10th) in the elastic motif are hydrophilic or hydrophobic respectively. Moreover, each pure silk-like protein was able to spontaneously self-assemble into films from aqueous solutions. Two kinds of synthetic fibers were made by pulling fibers from these preassembled films as well as spinning fibers from each protein resolubilized in HFIP. The mechanical data show that the pulled fibers are far tougher than the spun fibers suggesting a better fiber organization.
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References
Gosline J, Denny M, DeMont M (1984) Nature 309:551
Stauffer S, Coguill S, Lewis R (1994) J Arachnol 22:5
Gosline J, Guerette P, Ortlepp C, Savage K (1999) J Exp Biol 202:3295
Denny M (1976) J Exp Biol 65:483
Hinman M, Lewis R (1992) J Biol Chem 267:19320
Hayashi C, Shipley N, Lewis R (1999) Int J Biol Macromol 24:271
Simmons A, Michal C, Jelinski L (1996) Science 271:84
Termonia Y (1994) Macromolecules 27:7378
Hayashi C, Lewis R (1998) Mol Biol 275:773
Hayashi C, Lewis R (2000) Science 287:1477
Becker N, Oroudjev E, Mutz S, Cleveland J, Hansma P, Hayashi C, Makarov D, Hansma H (2003) Nat Mater 2:278
Hayashi C, Lewis R (2001) BioEssays 23:750
Zhou Y, Wu S, Conticello V (2001) Biomacromolecules 2:111
Chang D, Ventachalam C, Prasad K, Urry D (1989) J Biomol Struct Dynamics 6(i5):851
Urry D, Shaw R, Prasad K (1985) Biochem Biophys Res Commun 130(i1):50
Urry D, Hugel T, Seitz M, Gaub H, Sheiba L, Dea J, Xu J, Parker T (2002) Phylos Trans R Soc London, Ser B 357(i1418):169
Tatham A, Hayes L, Shewry P, Urry D (2001) Biochim Biophys Acta 1548:187
Hutchinson E, Thornton J (1994) Protein Sci 3:2207
Wilmot C, Thornton J (1988) J Mol Biol 5:221
Lewis R, Hinman M, Kothakota S, Fournier M (1996) Protein Expr Purif 7:400
Birnboim H, Doly J (1979) Nucleic Acids Res 7:1513
Sambrook J, Fritsch E, Maniatis T (1989) Molecular cloning. A laboratory manual. Second Edn. Cold Spring Harbor Laboratory
Wong C, Sridhara S, Bardwell J, Jakob U (2000) BioTechniques 28:426
Whithmore L, Wallace B (2004) Nucleic Acids Res, Web Server issue 24:W668
Scheller J, Guhrs K, Grosse F, Conrad U (2001) Nature 19:573
Teulé F, Jung S, Wood J, Marcotte W, Ellison M, Abbott A (2002) In: Brebbia C, Sucharov L, Pascolo P (eds) Design and nature, Udine, Italy. WIT press, p 379
Prince J, McGrath K, DiGirolamo C, Kaplan D (1995) Biochemistry 34:10879
Fahnestock S, Irvin S (1997) Appl Microbiol Biotechnol 47:23
Fahnestock S, Bedzyk L (1997) Appl Microbiol Biotechnol 47:33
Arcidiacono S, Mello C, Kaplan D, Cheley S, Bayley H (1998) Appl Microbiol Biotechnol 49:31
Dicko C, Knight D, Kenney J, Vollrath F (2004) Biomacromolecules 5:2105
Vollrath F, Knight D (2001) Nature 410:541
Knight D, Vollrath F (2001) Naturwissenschaften 88:179
Robson P, Wright G, Sitarz E, Maiti A, Rawa M, Youson J, Keeley F (1993) J Biol Chem 268(2):1440
Rauscher S, Baud S, Miao M, Keeley F, Pomès R (2006) Structure 14:1667
Creighton T (1993) Proteins: structures and molecular properties, Second Edn. Freeman, New York
Haq S, Khan R (2005) Int J Biol Biomacromolecules 36:47
Sponner A, Schlott B, Vollrath F, Unger E, Grosse F, Weisshart K (2005) Biochemistry 44:4727
Xu M, Lewis R (1990) Proc Natl Acad Sci USA 87:7120
Chen X, Shao Z, Vollrath F (2006) Soft Matter 2:448
Blackledge T, Hayashi C (2006) J Exp Biol 209:2452
Blackledge T, Hayashi C (2006) J Exp Biol 209:3131
Acknowledgements
We would like to thank Dr Michael B Hinman (Department of Molecular Biology, University of Wyoming) for providing the original A1, Y1 and S8 pBluescript clones, and for performing the amino acid analyses of the recombinant proteins. We also want to thank Professor Michael S Ellison (School of Materials Science and Engineering, Clemson University) for his valuable advice on the mechanical testing methods and analyses. This research was supported by grants from NIH, NSF and AFoSR.
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Teulé, F., Furin, W.A., Cooper, A.R. et al. Modifications of spider silk sequences in an attempt to control the mechanical properties of the synthetic fibers. J Mater Sci 42, 8974–8985 (2007). https://doi.org/10.1007/s10853-007-1642-6
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DOI: https://doi.org/10.1007/s10853-007-1642-6