Research Proposal: Angela Belcher CHEM*7530. Scott Allen Physics Department Winter [Mao, Science, 2004]

Transcription

1 Research Proposal: Angela Belcher [Mao, Science, 2004] Scott Allen Physics Department Winter 2006 CHEM*7530

2 Outline Background introduce Angela Belcher display technology research performed in the Belcher group the Proposal optical manipulation the engineered virus the goal Summary

3 Angela Belcher, Ph.D. BA, Creative Studies, University of California, Santa Barbara, 1991 PhD, Chemistry, University of California, Santa Barbara, 1997 Professor of Materials Science and Engineering and Biological M.I.T. Du Pont Young Investigators Award (1999) Presidential Early Career Award in Science and Engineering (2000) MacArthur Fellowship Genius Grant (2004)

4 Belcher Group uses nature as a guide to develop novel electronic and magnetic materials and to pattern materials on the nanoscale pioneered a very novel, non-covalent, self-organizing approach utilizing evolutionary peptide selection and peptide engineering to specifically recognize, bind, and grow nanoscale building blocks using selected peptides they have successfully controlled the physical properties of nanocrystals and subsequently employed molecular recognition and self-assembly in the design of hybrid biological-inorganic materials.

5 Display technology certain biological molecules exist that display amino acid sequences or peptides to their environment DNA manipulation allows for the systematic variation of the peptides that are on display a library of the display variants can be created several display platforms currently in use are bacteria [1-5], phage [6-17] and yeasts [18,19]

6 Using a display library target materials can be referenced against the library to discover peptide sequences that have specific affinity for the target Left with molecules having specific affinity for the target TARGET

7 Phage display early work in the Belcher lab focused on the M13 phage (virus) as its main display technology 880 nm DNA container 6.5 nm **not to scale encapsulated with roughly 2800 copies of the major coat protein VIII (pviii) capped at one end with approximately five molecules each of pvii and pix other end has about five molecules each of piii and pvi

8 Belcher group research highlight [6] Whaley et al., Nature, 2000 Selection of peptides with semiconductor binding specificity for directed nanocrystal assembly piii coat protein engineered combinatorial library created ~ amino acid peptide combinations experimented with five different single-crystal semiconductors: GaAs(100), GaAs(111)A, GaAs(111)B, InP(100) and Si(100) identified selective peptide binding: crystal composition binding to GaAs but not to Si crystalline face binding to (100) GaAs but not (111)B GaAs

9 Belcher group research highlight [7] Lee et al., Science, 2002 Ordering of Quantum Dots Using Genetically Engineered Viruses able to self assemble 3D structures created viral films containing ZnS nanocrystals

10 Belcher group research highlight [8] Lee et al., Adv. Mater., 2003 Virus-Based Alignment of Inorganic, Organic, and Biological Nanosized Materials any molecule covalently linked to streptavidin can be bound by M13

11 Belcher group research highlight [15] Nam et al., Nano Lett., 2004 Genetically Driven Assembly of Nanorings Based on the M13 Virus bifunctional M13 viruses displayed an anti-streptavidin peptide (piii) and hexahistidine peptide (pix) at opposite ends of the virus his peptide Ni-NTA Linker Molecule strep anti-strep

12 Belcher group research highlight [16] Huang et al., Nano Lett., 2005 Programmable Assembly of Nanoarchitectures Using Genetically Engineered Viruses bifunctional viruses displayed an antistreptavidin peptide (piii) and a selected gold binding peptide (pviii) TEM images C streptavidin conjugated CdSe (15 nm) A,B,D,E streptavidin conjugated Au (15 nm) ~ 5 nm Au bound to viral body

13 Belcher group research highlight [18] Peele et al., Acta Biomater., 2005 Probing the interface between biomolecules and inorganic materials using yeast surface display and genetic engineering [19] Peele et al., Langmuir, 2005 Design Criteria for Engineering Inorganic Material-Specific Peptides first attempts at developing a fundamental understanding of the interaction between the displayed peptides and the inorganic surfaces (1) Which amino acid functional groups are sufficient for binding short peptides to the inorganic material surfaces? (2) How do neighboring amino acid functional groups and their spatial arrangement in a peptide sequence modulate binding strength? (3) Can our results be used to predictively design peptides specific for these different material surfaces?

14 Belcher group research highlight used yeast as the display platform 4 mm spherical cells that are easily visualized by conventional light microscopy created a library of yeast cells displaying ~10 9 different peptide sequences looked at binding to CdS, CdSe, ZnS, and ZnSe and Au surfaces gives relative binding affinities for each displayed peptide

15 Belcher group research highlight through systematic genetic engineering of the specific peptide sequence displayed the Belcher group concluded that: only histidine, tryptophan, methionine, and cysteine appeared sufficient for significant binding spatially proximal amino acids could be altered to tune the binding strength

16 Proposal undoubtedly a fantastic collection of research BUT binding studies are semi-quantitative at best further research will focus on: a quantitative analysis to elucidate the forces involved in the binding of displayed peptide sequences to inorganic surfaces the effect of environmental conditions (ph, temperature, ionic species, ionic concentration) on binding affinity the creation of a novel and generic quantitative binding assay platform

17 Bustamante, Nat. Rev. Mol. Cell Biol., 2000 Proposal optical manipulation can control single nanoparticles with angstrom precision and measure interaction forces in the sub-pico-newton regime [20,21] movement away from the focus of the laser induces a restoring force that is proportional to the distance moved able to probe behaviour of materials in isolation traditional experimental techniques offer ensemble average responses

18 Proposal optical manipulation [22] Hansen et al., Nano Lett., 2005 Expanding the Optical Trapping Range of Gold Nanoparticles recently single-beam optical trapping in three dimensions of gold spheres with diameters ranging from 18 to 254 nm has been demonstrated pix piii gold binding motif peptide using past experience engineered M13 could be created to bind Au nanoparticles at pix thereby enabling a handle for optical manipulation

19 Proposal displacement from trap centre enables the study of a single M13 virus instead of yeast bring surface toward M13 virus allowing piii time to bind pull surface away from optical trap study binding force

20 Proposal the goal this platform will enable the systematic quantitative study of binding forces between displayed peptides and inorganic surfaces environmental conditions can easily be altered to study effect on binding affinity ph, temperature, ionic species, ionic concentration the creation of a novel and generic quantitative binding assay can be created via molecules conjugated to streptavidin pix piii gold binding motif anti-streptavidin

21 Summary described past work done in the lab of Angela MIT introduced display technologies in particular M13 phage display peptide display libraries can be used to bind and order inorganic nanocrystals also any molecule conjugated to streptavidin Belcher lab made efforts to elucidate fundamentals of peptide-surface binding through use of yeast display semi-quantitative presented proposal for quantitative analysis of peptide-surface binding forces optical manipulation and genetic engineering of M13

22 References [1] S. Brown, Proc. Natl. Acad. Sci. U.S.A., 89, 8651 (1992). [2] S. Brown, Nature Biotechnol., 15, 269 (1997). [3] S. Brown, M. Sarikaya and E. Johnson, J. Mol. Biol., 299, 725 (2000). [4] M.A. Schembri, K. Kjaergaard and P. Klemm, FEMS Microbiol. Lett., 170, 363 (1999). [5] K. Kjaergaard, J.K. Sorensen, M.A. Schembri and P. Klemm, Appl. Environ.Microbiol., 66, 10 (2000). [6] S.R. Whaley, D.S. English, E.L. Hu, P.F. Barbara and A.M. Belcher, Nature, 405, 665 (2000). [7] S.W. Lee, C. Mao, C.E. Flynn and A.M. Belcher, Science, 296, 892 (2002). [8] S.W. Lee, S.K. Lee and A.M. Belcher, Adv. Mater., 15, 689, (2003). [9] R.R. Naik, S.J. Stringer, G. Agarwal, S.E. Jones and M.O. Stone, Nature Mater., 1, 169 (2002). [10] C. Mao, C.E. Flynn, A. Hayhurst, R.Y. Sweeney, J. Qi, G. Georgiou, B. Iverson and A.M. Belcher, Proc. Natl. Acad. Sci. USA, 100, 6946 (2003). [11] S. Wang, E.S. Humphreys, S.-Y. Chung, D.F. Delduco, S.R. Lustig, H. Wang, K.N. Parker, N.W. Rizzo, S. Subramoney, Y.-M. Chiang and A. Jagota, Nature Mater., 2, 196 (2003). [12] C.E. Flynn, C. Mao, A. Hayhurst, J.L. Williams, G. Georgiou, B. Iverson and A.M. Belcher, J. Mater. Chem., 13, 2414 (2003).

23 References [13] S.-W. Lee and A.M. Belcher, Nano Letters, 4, 387 (2004). [14] C. Mao, D.J. Solis, B.D. Reiss, S.T. Kottmann, R.Y. Sweeney, A. Hayhurst, G. Georgiou, B. Iverson and A.M. Belcher, Science, 303, 213 (2004). [15] K.T. Nam, B.R. Peelle, S.-W. Lee and A.M. Belcher, Nano Letters, 4, 23 (2004). [16] Y. Huang, C.-Y. Chiang, S.K. Lee, Y. Gao, E.L. Hu, J. De Yoreo and A.M. Belcher, Nano Letters, 5, 1429 (2005). [17] S.-K. Lee, D.S. Yun and A.M. Belcher, Biomacromolecules, 7,14 (2006). [18] B.R. Peelle, E.M. Krauland, K.D. Wittrup and A.M. Belcher, Acta. Biomater., 1, 145 (2005). [19] B.R. Peelle, E.M. Krauland, K.D. Wittrup and A.M. Belcher, Langmuir, 21, 6929 (2005). [20] K. Dholakia and P. Reece, nanotoday, 1, 18 (2006). [21] C.A. Mirkin, R.L. Letsinger, R.C. Mucic and J.J. Storhoff, Nature, 382, 607 (1996). [22] P.M. Hansen, V.K. Bhatia, N. Harrit and L. Oddershede, Nano Letters, 5, 1937 (2005). [23] M. Rief, F. Oesterhelt, B. Heymann and H.E. Gaub, Science, 275, 1295 (1997). [24] J. Ni, S.-W. Lee, J.M. White and A.M. Belcher, J. Poly. Sci. B, 42, 629 (2004). [25] C. Bustamante, J.C. Macosko and G.J.L.Wuite, Nat. Rev. Mol. Cell Biol., 1, 130 (2000).

24 Extra Slides

25 Fibre spinning

26 Belcher group research highlight [13] Lee et al., Nano Lett., 2004 Virus-Based Fabrication of Micro- and Nanofibers Using Electrospinning liquid crystalline M13 viruses extruded from a capillary tube into a crosslinking solution of glutaraldehyde fibers were mm in diameter M13 viruses were mixed with polyvinyl pyrolidone (PVP) and electrospun into nanofibres virus-pvp electrospun fibers maintained their ability to infect bacterial hosts after resuspending in buffer solution

27 Force Spectroscopy very similar to force spectroscopy done using AFM Single Molecule Force Spectroscopy on Polysaccharides by Atomic Force Microscopy [23] Rief et al., Science, 1997