Functional nanotubes
200((will be filled in by the editorial staff))
Peptide Nanotubes as Light Harvesting Antenna**
Yan Liang, Peng Guo, Sai Venkatesh Pingali, Suzette Pabit, Pappannan Thiyagarajan*, Keith M. Berland*, and David G. Lynn*
The reaction center and light-harvesting complexes are remarkable
for their precise pigment arrangement and long range order.1-6
Artificial light harvesting complexes and reaction centers have
attempted to use the natural ordering of isolated bacterial protein-
membrane fragments7-9 within lipid bilayer liposomes,4,10-12 or
polymeric assemblies to order pigments.13-15 Here we build on
paracrystalline amyloid assembly16 to construct pigment arrays on a
new peptide nanotube scaffold and exploit Förster resonance energy
transfer 17 measurements to probe the resulting functionalized surfaces.
a)
b)
Figure 1. Nanotube formed by Aβ(16-22) at pH 2, (a) TEM image,
scale=100nm; (b) the proposed model and diameters of the nanotubes.
16,18
The inset shows the Aβ(16-22) bilayer model for the nanotube shell.
[∗] Y. Liang, Prof. Dr. D. G. Lynn
Center for the Analysis of Supramolecular Self-assemblies,
Departments of Chemistry and Biology, Emory University
Atlanta, GA 30322 (USA)
Fax: (404) 727-6586
E-mail: dlynn2@emory.edu
P. Guo, Dr. Suzette Pabit, Prof. Dr. K. M. Berland
Department of Physics, Emory University
Atlanta, GA 30322 (USA)
E-mail: keith.berland@emory.edu
Dr. S. V. Pingali, Dr. P. Thiyagarajan
Advanced Photon Source, Argonne National Laboratory
Argonne, Illinois 60439 (USA)
E-mail: thiyaga@anl.gov
[∗∗] This work benefited from the Apkarian Microscopy Center at
EU, assistance from Dr. Soenke Seifert at the APS under
U.S. DOE contracts BES and W-31-109-ENG-38 through the
University of Chicago, support from DOE (ER15377), NSF
(CRC-CHE-0404677), and NSF (CBC-CHE-0739189), and
CD instrumentation from NSF(CHE-0131013).
Supporting information for this article is available on the
WWW under http://www.angewandte.org or from the author.
DOI: 10.1002/anie.
Aβ(16-22), the seven-residue amyloid-β segment Ac-KLVFFAE-
NH2, self-assembles into soluble amyloid nanotubes at pH 2 (Fig.
1a).17 These Aβ(16-22) nanotubes maintain antiparallel one-residue
shifted β-sheet bilayers (Fig. 1b) and the cross-β architecture,18
creating a nanotube surface that positions the peptide termini in a
repeating 5Å X 10Å rectangular pattern. Amyloid fibrils display
properties useful for functional molecular engineering,19-20,22-23 and
this patterned array of the nanotubes appeared to be suitable for light
harvesting opportunities.
The N- and C-terminal capped Aβ(16-22) at pH 2 contains a
hydrophobic core, LVFFA, buttressed by polar Lys and Glu residues
at the N- and C-termini. N-terminal capping with rhodamine (Rh110)
introduces an additional positive charge, increases solubility in
CH3CN:H2O (2:5, v:v), and the resulting Rh16-22 readily forms
amyloid fibrils (Fig. S1). Fourier transform infrared (FTIR) analysis
confirmed an amide I band shift from 1639cm-1 to 1627cm-1 (Fig. 3a),
very similar to Aβ(16-22),18 but with an additional rhodamine
signature at 1597cm-1. Wide angle X-ray scattering (WAXS)19 further
established the amyloid cross-β diffraction pattern with 5Å (H-
bonding) and broad 10Å (lamination) bands (Fig. S2), confirming the
rhodamine containing peptide assembly maintains the cross-β pattern
of amyloid.
When Aβ(16-22) and Rh16-22 (250:1 molar ratio) are allowed to
co-assemble as a 1mM peptide solution in CH3CN:H2O (2:3, v:v) at
pH 2, the resulting nanotubes are readily observed by transmission
electron microscopy (TEM) and two-photon fluorescence imaging
(Fig. 3). The morphologies of these assemblies are identical to the
Aβ(16-22) nanotubes by TEM and the fluorescence appears
homogeneously distributed across each nanotube. Rh110 alone, Fig.
3b, does not bind to the Aβ(16-22) nanotubes, underscoring the
requirement for peptide co-assembly.
1627 12
a C, 1640 b
13
C, 1599
1597
1639
1610
1597
1800 1700 1600 1500 1800 1700 1600 1500
-1 -1
Wave number (cm ) Wave number (cm )
Figure 2 a. FTIR of Aβ(16-22) nanotubes (black), Rh16-22 fibers
13
(green), and unassembled Rh16-22 (red). b. IE-FTIR of [1- C-F19]
13
Aβ(16-22) (blue), Aβ(16-22) + [1- C-F19] Rh16-22 (10:1) (red), Aβ(16-
22) + Rh16-22 (10:1) (green), and Aβ(16-22) (black).
The introduction of [1-13C] labels into β-strands splits the amide I
band through coupling of the 12C and 13C dipoles within the sheet.24-26
For example, isotope-edited FTIR (IE-FTIR) of [1-13C]-F19
incorporated Aβ(16-22) splits the main amide I band into components,
1640 cm-1 and 1599 cm-1 (Fig. 2b).18 With [1-13C]-F19 Rh16-22 at
0.15mM, well below its critical assembly concentration, co-assembly
with 1.5mM Aβ(16-22) gives a distinct red-shifted shoulder on the
main 1627cm-1 amide I band (Fig. 2b). This band is absent in the
1
 a Rh16-22 b Rh110 c. A555

Figure 3 The top panels are two-photon fluorescence imaging of nanotubes, and the bottom panels are corresponding TEM images. a.
nanotubes formed by Aβ(16-22):Rh16-22 co-assembly, (250:1 molar ratio); b. Aβ(16-22) mature nanotubes with Rh110, (250:1 molar ratio); c.
Aβ(16-22) mature nanotubes with A555 (1000:1 molar ratio). Fluorescence scale=5µm, TEM scale=100nm.

To probe the pattern accessible to the nanotube surface, we explored

a) b) other binding molecules. Congo red staining is diagnostic of amyloid,
and when bound to nanotubes,16,18 shows a characteristic birefringence
under polarized light.21 Alexa 555 (A555) has a similar structure, with
sulfate functionality on a planar aromatic nucleus. When mixed with
mature Aβ(16-22) assemblies, the A555 dye decorates the nanotubes,
immediately forming fluorescent tubes (Fig. 2d), consistent with its
ability to bind to the surface of the nanotubes. The Rhodamine 110 and
A555 dyes are a FRET donor/acceptor pair (Fig S3), and when A555
is added to mature co-assembled Aβ(16-22)/Rh16-22 nanotubes
0 1000 2000 3000 4000 (A555/Rh16-22/Aβ(16-22) 1:4:1000 with Aβ(16-22) at 0.5mM), the
Time / ps center of the Rh16-22 lifetime distribution shifts from 3.7 to 3.3ns
c)
(Fig. 4b). This corresponds to a FRET efficiency of 11%, calculated as
1-τ’/τ, where τ’ is the Rh16-22 lifetime in the presence of A555, and τ
is the lifetime in its absence.27 The FRET signal indicates that the two
10nm
different dyes are positioned in close proximity along the nanotubes.
For every FRET donor and acceptor pair, the energy transfer
efficiency is inversely proportional to the sixth power of their
separation. While it is difficult to precisely estimate the Rh16-22 and
A555 distribution within the nanotubes, given the initial concentration
ratio and assuming minimal impact of the attached chromophore on
Figure 4 FRET lifetime analysis. a. Lifetime Image of Aβ(16-22):Rh16- peptide incorporation frequency, we estimate every 10 laminates
22 fluorescence nanotubes, scale=5μm; b. Lifetime distribution of A555 (10nm) of 12 β-strands (12.5nm) should have one Rh16-22 peptide, as
(red) with mature Aβ(16-22) nanotubes and Aβ(16-22):Rh16-22 represented by the green balls in Fig. 4c, right panel. When this
fluorescence nanotubes in the presence (black) and absence (blue) of nanotube pattern is further layered with A555 at a 4X lower
A555; c. left: schematic of 2D distribution of Rh16-22 in Aβ(16-22) concentration (the red balls), the approximate spacing between the
bilayer, and the model of FRET between Rh16-22 and A555, blue: the donor and acceptor is estimated on the order of 10nm. The observed
peptide bilayer, red: A555, green: Rh110 in Rh16-22; right: the ratio of 11% FRET efficiency is consistent with this spacing given a Förster
Rh16-22 and A555 in Aβ(16-22) bilayer based on the concentration radius R0 of 6.6 nm for Rh16-22 and A555 (Fig. S3)
ratio. These results are most consistent with the functionalized Rh16-22
peptides being randomly incorporated as the Aβ(16-22) nanotubes
assemble. This ability to assemble large chromophores across the
assembly lacking the 13C label, providing support for co-assembly of paracrystalline 2D surface of hollow peptide nanotubes offers a
the Rh16-22 peptide within the Aβ(16-22) β-sheets. network that is ordered, like lipid membranes, along the well defined

2
inner and outer compartments of the nanotube. Moreover, given the
dimensions of the array, it should now be possible to incorporate
further molecular recognition elements, construct higher order
arrays,28 and include elements for energy and electron separation
reactions. Accordingly, this extension of amyloid self-assembly to
more precise supramolecular arrays containing functional pigments
provides a critical scaffold for constructing new bio-inspired antenna
and photosynthetic devices.

Received: ((will be filled in by the editorial staff))

Published online on ((will be filled in by the editorial staff))

Keywords: amyloid · FRET· light harvesting · peptides nanotubes · self-

assembly

[1] Allen, J. P.; Feher, G.; Yeates, T. O.; Komiya, H.; Rees, D. C., Proc. Natl.
Acad. Sci. USA 1987, 84, 5730-5734.
[2] Allen, J. P.; Williams, J. C., FEBS Lett. 1998, 438 5-9.
[3] Damjanovic, A.; Kosztin, I.; Kleinekathofer, U.; Schulten, K., Phys. Rev. E
2002, 65, 31919-31953.
[4] Jones, M. R., Prog. Lipid Res. 2007, 46, 56-87.
[5] Deisenhofer, J.; Epp, O.; Miki, K.; Huber, R.; Michel, H., J. Mol. Biol. 1984,
180, 385-398.
[6] Chang, C.-H.; El-Kabbani, O.; Tiede, D.; Norris, J.; Schiffer, M.,
Biochemistry 1991, 30, 5352-5360.
[7] Goca, J.; Harab, M.; Tateishia, T.; Miyake, J., J. Photochem. Photobiol. A:
Chemistry 1996, 93, 137-144.
[8] Iida, K.; Kiriyama, H.; Fukai, A.; Konings, W. N.; Nango, M., Langmuir
2001, 17 2821-2827.
[9] Dewa, T., Selective e-Journal of Surface Science and Nanotechnology 2005
3, 145-150.
[10] Páli, T.; Garab, G.; Horváth, L. I.; Kóta, Z., Cell. Mol. Life Sci. 2003, 60,
1591-1606.
[11] Bahatyrova, S.; Frese, R. N.; Siebert, C. A.; Olsen, J. D.; Werf, K. O. v. d.;
Grondelle, R. v.; Niederman, R. A.; Bullough, P. A.; Otto, C.; Hunter, N.,
Nature 2004, 430, 1058-1062.
[12] Scheuring, S.; Sturgis, J. N.; Prima, V.; Bernadac, A.; Levy, D.; Rigaud,
J.-L., Proc. Natl. Acad. Sci. USA 2004, 101, 11293–11297.
[13] Webber, S. E., Chem. Rev. 1990, 90, 1469-1482.
[14] Watkins, D. M.; Fox, M. A., Desk Reference of Functional Polymers:
Syntheses and Applications 1997, Chapter 4, 549-566.
[15] Hasobe, T.; Saito, K.; Kamat, P. V.; Troiani, V.; Qiu, H.; Solladie, N.;
Kim, K. S.; Park, J. K.; Kim, D.; D’Souza, F.; Fukuzumi, S., J. Materials
Chem. 2007, 17, 4160–4170.
[16] Lu, K.; Jacob, J.; Thiyagarajan, P.; Conticello, V. P.; Lynn, D. G., J. Am.
Chem. Soc. 2003, 125, 6391-6393.
[17] a) B. W. v. d. Meer, Resonance energy transfer : theory and data, New
York : VCH, 1994. b) P. K. Wolber, B. S. Hudson, Biophys. J. 1979, 28,
197. c) K. B. Towles, A. C. Brown, S. P. Wrenn, N. Dan, Biophys. J.l
2007, 93, 655–667. d) S. V. Solomatin, T. K. Bronich, A. Eisenberg, V.
A. Kabanov, A. V. Kabanov, Langmuir 2007, 23, 2838.
[18] Mehta, A. K.; Lu, K.; Childers, W. S.; Liang, Y.; Dublin, S.; Dong, J.;
Snyder, J. S.; Pingali, S. V.; Thiyagarajan, P.; Lynn, D. G., J. Am. Chem.
Soc. 2008, (in press)
[19] Gilead, S.; Gazit, E., Supermol. Chem. 2005, 17, 87-92.
[20] Hamada, D.; Yanagihara, I.; Tsumoto, K., TRENDS Biotechnol. 2004, 22,
93-97.
[21] Elghetany, M. T.; Saleem, A.; Barr, K., Ann.Clin.Lab. Sci., 1989, 19, 190-
195.
[22] MacPhee, C. E.; Dobson, C. M., J. Am. Chem. Soc. 2000, 122, 12707-
12713.
[23] Kodama, H.; Matsumura, S.; Yamashitaa, T.; Mihara, H., Chem. Commun.
2004, 2876-2877.
[24] Hiramatsu, H.; Kitagawa, T., Biochim. Biophys.Acta 2005 1753, 100 – 107.
[25] Petty, S. A.; Decatur, S. M., J. Am. Chem. Soc. 2005, 127, 13488-13489.
[26] Paul, C.; Axelsen, P. H., J. Am. Chem. Soc. 2005, 127, 5754-5755.
[27] Wallrabe, H.; Periasamy, A., Curr. Opin. Biotechnol., 2005, 16, 19-27.
[28] Lu, K.; Guo, L.; Mehta, A.K.; Childers, W.S.; Dublin, S.N.; Conticello,
V.P.; Thiyagarajan, P.; Apkarian, R.P.; Lynn, D.G. 2007 Chem Comm.
2729 – 2731.

3
Entry for the Table of Contents

Functional Nanotubes

Yan Liang, Peng Guo , Sai Venkatesh nanotube

Pingali, Suzette Pabit, Pappannan functionalization
Thiyagarajan, Keith M. Berland, and David G.
Lynn*

Peptide Nanotubes as Light Harvesting

Antenna
Light harvesting antenna: a 2D pigment array is constructed within Aβ(16-22)
nanotubes. The incorporated pigment array is capable of Förster energy transfer,
establishing a starting point for constructing new bio-inspired antenna and
photosynthetic devices.