Building Nanowires from Micelles: Hierarchical Self-Assembly of Alternating Amphiphilic Glycopolypeptide Brushes with Pendants of High-Mannose Glycodendron and Oligophenylalanine

Article abstract of DOI:10.1021/jacs.6b05044

Mimicking the diverse glyco-conjugate structures in nature is always the dream of scientists. Right now, hierarchical self-assembled structures of natural conjugates of peptides and sugars could not easily be achieved via linear glycopolypeptide with mono

Full text of DOI:10.1021/jacs.6b05044

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Article
Building Nanowires from Micelles: Hierarchical Self-assembly
of Al-ternating Amphiphilic Glycopolypeptide Brushes with
Pendants of High-Mannose Glycodendron and Oligophenylalanine
Yijiang Liu, Yufei Zhang, Zheyu Wang, Jue Wang, Kongchang Wei, Guosong Chen, and Ming Jiang
J. Am. Chem. Soc., Just Accepted Manuscript • DOI: 10.1021/jacs.6b05044 • Publication Date (Web): 22 Jul 2016
Downloaded from http://pubs.acs.org on July 23, 2016
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Page 1 of 17
Journal of the American Chemical Society
Building Nanowires from Micelles: Hierarchical Self-assembly of Al-
ternating Amphiphilic Glycopolypeptide Brushes with Pendants of
High-Mannose Glycodendron and Oligophenylalanine
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Yijiang Liu, Yufei Zhang, Zheyu Wang, Jue Wang, Kongchang Wei, Guosong Chen*, Ming Jiang
The State Key Laboratory of Molecular Engineering of Polymers and Department of Macromolecular Science, Fudan Uniꢀ
versity, Shanghai 200433, China
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ABSTRACT: Mimicking the diverse glycoꢀconjugate structures in nature is always the dream of scientists. Right now, hierarchical
selfꢀassembled structures of natural conjugates of peptides and sugars could not easily be achieved via linear glycopolypeptide with
monosaccharides as attachments. In this work, by using a series of wellꢀdesigned alternating amphiphilic glycopolypeptide brushes
(
AAGBs) with pendants of glycodendrons and short peptides, various selfꢀassembled morphologies were achieved, including nanꢀ
owires, nanoribbon and compound micelles mainly depending on the number ratio of the sugar units to the amino acids species
S/F). Among these morphologies, nanowire attracted our great attention. TEM studies demonstrated that it is formed via a hierarꢀ
(
chical selfꢀassembly i.e. a series of successive processes, including micellization, micelles alignment forming nanoꢀfilament,
branching of the nanoꢀfilaments by micelles, and finally nanowire formation. As far as we know, such hierarchical selfꢀassembly
process with high complexity has not been observed in literature for glycopolypeptides even polypeptides, which will deepen our
understanding on selfꢀassembly mechanism of natural glycoꢀconjugates and expand the library of biomimetic materials.
tried a new type of glycopolypeptide, prepared by postꢀ
polymerization modification of a polypeptide backbone with
Introduction
oligosaccharide and oligopeptide as pendent brushes. The obꢀ
In nature, saccharides and peptides are often covalently conꢀ
tained new polymer is featured by the alternating amphiphilic
nected to each other forming various biomacromolar conjuꢀ
1
brushes of dendronized oligosaccharides and oligopeptides
along the polymeric peptide backbone. It is well known that
polymer brushes, particularly amphiphilic polymer brushes,
being a family of new architectures of copolymers have drawn
gates, such as glycoproteins, mucins and proteoglycans,
which can form a variety of assembled structures especially
2
hierarchical ones, including micelles, vesicles, wormꢀlike
3
micelles, nanowires, nanoribbons etc. These selfꢀassembled
23
great attention in the last two decades as their relatively well
defined structures provide broad opportunities to control hyꢀ
drophobicꢀhydrophilic balance and then the final assemblies.
However, in most cases their assemblies possess lamellar
structure as the result of phase separation of the side chains to
opposite sides of the polymer backbone. Some recent works
structures play key roles in a broad range of biological proꢀ
4
cesses including modulating intercellular communication, cell
5
6
adhesion, and immunological recognition. In the last decade,
7
significant progresses on bioinspired synthetic glycoꢀpeptides
have been achieved via different approaches aiming at mimꢀ
8
icking the functions of the native ones and developing new
2
4
25
9
introduce crystallizable chains, liquid crystal or bulky
biocompatible materials. Among these mimics, glycopolypepꢀ
26
1
0
POSS as brushes but did not bring out new findings in selfꢀ
assembly. It is also well known that short peptides are excelꢀ
lent building blocks to create a wide variety of nanostrucꢀ
tide prepared by ringꢀopening polymerization is a promising
11
one, featured by facile preparation and easy scaleꢀup. In this
field, artificial glycopolypeptides with a polypeptide backbone
and monosaccharides as pendent groups were prepared by
27
28
29
tures, some of which, such as nanowire, nanoribbon reꢀ
30,31
1
2
sembles those in nature.
However, such strong assembly
controlled polymerization, and in some cases, followed by
1
3
ability of peptide leading to morphologies found in nature has
rarely recurred in brush polymers with peptide as side chains,
postꢀpolymerization modification. In these artificial glycoꢀ
polypeptide systems, the carbohydrates not only contribute to
32
1
4
15
and even less in glycopolypeptides.
the bioavailability, but also modulate the folding and the
16
selfꢀassembly of the conjugates. However, among these eleꢀ
In our design, the glycodendron attachment makes the
conjugates unique in very high density of sugar units, of which
gant works in the past, very differing from the architectural
varieties of their native counterparts, the artificial glycopolyꢀ
peptides studied were mainly focusing on linear or linearꢀ
33
carbohydrateꢀcarbohydrate interaction may promote assemꢀ
bly of the conjugates; meanwhile, the pendent βꢀsheet forming
oligopeptide is expected to provide driving force for the asꢀ
sembly resemble to that occurring in nature. Thus, new glycoꢀ
polypeptide architecture, i.e. Alternating Amphiphilic Glycoꢀ
peptide Brushes (AAGBs) was prepared by modification of
Manα1ꢀ2Man oligomannosides and oligophenylalanines to the
1
7
branched block copolymers, which usually led to zero dimenꢀ
18
tional structures, e.g. micelles and vesicles exclusively. Thus
these limitations hamper our understanding on the selfꢀ
1
9
assembly mechanism of native glycoꢀconjugates, as well as
the development of functional biomaterials based on glycoꢀ
2
0
21
polypeptides for drug delivery, bioꢀimaging, tissue engiꢀ
neering etc.
34
22
Lysine/Glutamate based dendritic backbone. Manα1ꢀ2Man
was selected because this structure was highly expressed on
35
the surface of virus and bacteria, e.g., gp120, a major enveꢀ
Aiming at creating more abundant morphologies, espeꢀ
cially the hierarchical selfꢀassembled ones from artificial glyꢀ
copolypeptides approaching to those in nature, in this work we
lope protein of HIV, which could be recognized by human
36
monoclonal antibody (mAbs) 2G12.
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Scheme 1. Synthetic routes and chemical structures of dendritic polypeptide scaffolds P0, P1 and P2.
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Scheme 2. Synthesis and chemical structures of (a) Alternating Amphiphilic Glycopolypeptide Brush (AAGB) P2tM-F . (b)
Full space model and (c) 3D schematic illustration of the repeating unit of P2tM-F4.
4
Meanwhile, the oligophenylalanine (LꢀPhe , FF or FFFF) exꢀ
previously in any research on brush copolymers. Furthermore,
the protein binding ability of different assembled morpholoꢀ
gies was evaluated.
n
37
tracted from the Alzheimer’s βꢀamyloid polypeptide, was
found to have an excellent capacity in forming a wide range of
38
nanostructures. To prepare AAGBs, these two representative
components were linked alternatively as pendent groups onto a
polypeptide backbone prepared by condensation polymerizaꢀ
Result and Discussion
39
tion via our previous strategy.
Design and Synthetic Strategies of Alternating Amphiphilic
Glycopeptide Brushes (AAGBs). Very recently, we used metꢀ
alꢀfree stepꢀgrowth copolymerization to prepare hybrid copolꢀ
In order to explore the basic role of the complex amꢀ
phiphilic structure in controlling its assembly morphologies, a
small library of AAGBs has been constructed where the polyꢀ
meric structure was tuned by three parameters, i.e. generations
of the backbone dendrons, structure of the conjugated oligoꢀ
saccharides and number of the oligophenylalanine. We found
that the resulting AAGBs selfꢀassembled into various morꢀ
phologies, including compound micelle, nanowire, nanoribbon
etc., mainly depending on the ratio of mannoside number in
glycodendron and number of phenylalanine in oligophenylalaꢀ
nine component. Strong evidence showed that nanowires were
formed from hierarchical selfꢀassemble processes, i.e. the
AAGBs first formed micelles, the micelles then further
aligned into filaments, the filaments grew with branches via
micelles, and finally the filaments fused into nanowires. This
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9
ymers from oligopeptide blocks linked by small molecules .
In this paper, the same strategy has been employed. Briefly,
the stepꢀgrowth polymerization between pentafluorophenol
(PFp) active ester and primary amine functionalized monomer
was used due to their high reactive efficiency under mild conꢀ
ditions (Scheme 1). PFp active ester functionalized monomer
M0, M1, M2 and amineꢀfunctionalized monomer M-a based
on Lysine and Glutamate were first prepared separately via
liquid phase peptide synthesis (synthetic details in supporting
information, Scheme S1ꢀ5). Different generations of dendron
scaffolds were attached to PFp active ester monomer as penꢀ
dent groups. Then stepꢀgrowth polymerization was performed
by simply adding 1.2 eqv. N,NꢀDiisopropyl ethylamine
(DIPEA) to the equal molar mixture of M-a with M0, M1 or
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M2 in DMSO at room temperature. The resulted viscous
hierarchical selfꢀassembly mechanism has not been reported
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Figure 1. The TEM and AFM images of the nanowires formed by P1tM-F in water, (a) chemical structure, and 2D schematic illustration
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of P1tM-F , (b, c) low/highꢀmagnification TEM images, (d) cyroꢀTEM image and (e) AFM image of the P1tM-F nanowires.
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mixtures were allowed to precipitate in diethyl ether, giving
alkyneꢀfunctionalized precursors. These precursors were
deprotected in TFA and then modified with aminooxy monoꢀ
mer M-b, giving scaffolds P0, P1 and P2 (Scheme 1). The
aminooxy nucleophiles are introduced for their high reactivity
with reducing saccharides under mild reaction condition. Moꢀ
lar mass of P0, P1 and P2 was measured by gel permeation
chromatography with multiꢀangle light scattering detector
(GPCꢀMALS) in DMF. P0 and P1 gave the molar mass of
18.9 kDa and 16.6 kDa, respectively, while the molar mass of
P2 can hardly be measured due to its poor solubility in DMF
(molar mass of corresponding precursors were listed in Table
S1,dn/dc measurement and GPC traces were shown in Figure
S1ꢀ6).
polypeptides P0tM, P1tM and P2tM, the mannosylamine
1
content cannot be determined from H NMR due to signal
1
overlapping with other anomeric H signal and D O solvent
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residue. The molar mass of glycopolypeptides was measured
by gel permeation chromatography with multiꢀangle light scatꢀ
tering detector (GPCꢀMALS) in water (0.8% NaNO ) and
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listed in Table S1.
The alkyneꢀfunctionalized backbone provided active sites
for further postꢀpolymerization modification (Scheme 2). Azꢀ
ideꢀmodified oligophenylalanine M-c (F ) and M-d (F ) were
2 4
synthesized as the hydrophobic domain for their known high
tendency to form stable βꢀsheets. Modification of M-c or M-d
was performed by typical Cu(I) catalyzed CuAAC reaction (or
“
click”) in DMSO, resulting AAGBs. 5% of LiBr was added
Then reducing saccharides were modified to aminooxy
nucleophiles at the end of pendent dendron on scaffolds P0,
P1 and P2. The experiments were performed in acetate buffer
to increase the solubility of M-c/M-d. For example, after
clicking with M-d, P2tM was transformed to P2tM-F4.
(200 mM, pH 3.5) with 100 mM aniline as catalyst. Typically,
A Representative Self-Assembly of AAGB P1tM-F
. We
4
the scaffolds with aminooxy groups (P0, P1 and P2) were
dissolved in acetate buffer, after which 1.2 equivalent of corꢀ
responding saccharides (Scheme 2, M: Mannose; dM: Manα1ꢀ
2Man; tM: Manα1ꢀ2Manα1ꢀ2Man) were added separately and
the mixtures were kept at 40ºC. Relatively high conversion
ratios of the aminooxy groups were obtained (80ꢀ95%, calcuꢀ
first discuss the selfꢀassembly of AAGB P1tM-F with mediꢀ
4
um sizes of hydrophilic and hydrophobic domains (Figure 1a).
Typically, 2 mL water was added to 1 mL clear DMSO soluꢀ
tion of P1tM-F
dialysis against deionized water. Opalescence appeared sugꢀ
gesting formation of P1tM-F assemblies. After dialysis for 48
(30 mg/mL). Then the mixture was allowed to
4
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1
lated from H NMR, Table S2). Excess saccharides (2 equiv.
h, the solution was diluted to 2 mg/mL by adding deionized
water. A large amount of long nanowires was observed by
TEM (Figure 1b, c) with a uniform width around 25 nm. The
length of each nanowire was hardly determined due to the
entanglement of the wires, but obviously the length was at
least in a micrometer range. The nanowire morphology was
further confirmed by cryoꢀTEM (Figure 1d). And the image
under atomic force microscope (AFM) suggested that the
thickness of nanowire was around 10 nm (Figure 1e, Figure
S7ꢀ8). The nanowires inclined to entangle into large cluster at
or more) were also used for this reaction but no obvious inꢀ
crement of the conversion rate was observed. According to
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literature, carbohydrate oxime Oꢀethers were formed during
covalent coupling of reducing saccharides with aminooxy
nucleophiles. The reaction normally generates more oxime
rather than mannosylamine due to the higher chemical stability
of the former. Accordingly, 12~15% of the ringꢀclosed manꢀ
nosylamine were found among the aminooxy glycosylation
sites on the side chain (Table S2). For trimannosideꢀmodified
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Table 1. Sample codes, 2D illustrations, S/F ratios and the assembled morphologies of AAGBs library
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Sample codes
P0M-F2
Morphology
Precipitate
S/F
2D illustrations
Sample codes
P0M-F4
Morphology
Precipitate
S/F
2D illustrations
0.5
0.25
P1M-F2
P2M-F2
Precipitate
Nanowire
1.0
2.0
1.0
2.0
4.0
1.5
3.0
6.0
P1M-F4
P2M-F4
Precipitate
Precipitate
Precipitate
Nanoribbon
Nanowire
Precipitate
Nanowire
Nanowire
0.5
1.0
0.5
1.0
2.0
0.75
1.5
3.0
0
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0
P0dM-F2
P1dM-F2
P2dM-F2
P0tM-F2
P1tM-F2
P2tM-F2
Precipitate
P0dM-F4
P1dM-F4
P2dM-F4
P0tM-F4
P1tM-F4
P2tM-F4
Nanowire
Compound Micelle
Nanowire
Compound Micelle
Compound Micelle
ymers and polymeric brushes, which deserved further investiꢀ
gation in detail.
Self-Assembly of AAGB Library. To further explore the
general rule of AAGB assembly with emphasis on the effects
of glycodendron and oligophenylalanine brushes, a library of
AAGBs was designed varying in dendriticꢀpolypeptide scafꢀ
fold (P0, P1 and P2), hydrophobic peptide domain (F /F ) and
2
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oligomannoside domain (M, dM and tM, Table 1). The same
procedures as the mentioned for P1tM-F were used for the
4
sample preparation and their assembly.
Generally, during the process of dialysis where the mediꢀ
um turned from hydrophobic to hydrophilic, the 18 samples
showed very different responses: a great part of them, i.e. sevꢀ
en samples precipitated without any assembled structures.
Three samples showed compound micelles, P2dM-F , P1tM-
2
F and P2tM-F (Figure 2a, S9bꢀc). The nanowires, which
2
2
were seldom found in synthetic copolymers and interested us
most, became the common features of the samples including
P1dM-F , P2tM-F , P2M-F , P0tM-F and P2dM-F (Figure
2
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2
2
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bꢀc, S9dꢀf). Nanoribbon is a special case, obtained only from
Figure 2. TEM images of (a) P2dM-F (compound micelle), (b)
2
P1dM-F (Figure 2d). Different from the long and homogeneꢀ
4
P1dM-F (nanowire), (c) P2tM-F (nanowire) and (d) P1dM-F
2
4
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ous nanowire, the nanoribbon was heterogeneous with its
width distributed in the range of 50~100 nm. The ribbons were
not stable and easy to precipitate in water. At first glance, such
diversity of the morphologies observed seems not easy to be
explained by relating the results to the structural parameters
directly.
(
nanoribbon).
high concentration ([P1tM-F ] >1 mg/mL) in less than 7 days
Figure S9a), but they are quite stable for more than 30 days at
a low concentration ([P1tM-F ] < 0.2 mg/mL) at room temꢀ
perature. We noticed that such long and uniform nanowires are
rarely observed for the assemblies of amphiphilic block copolꢀ
4
(
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slower assembly process and therefore, it was selected for this
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mechanical study. In addition, to further retard the process, a
highly diluted solution (2 mg/mL) of P2tM-F was used for
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dialysis. Thus the assembly process finished in about 2 days,
which made the TEM sample preparation at different stages
possible. Typically, at each time intervals of 3 h, 12 h, 24 h
and 48 h, a small aliquot of the solution was taken out and
quenched by a large amount of water to "freeze" the intermeꢀ
diate morphology.
The TEM image of P2tM-F taken at 3 h (Figure 4a) was
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featured by the coexistence of micelles with a diameter around
3
ꢀ5 nm and nanoꢀfilaments with the same width. It could be
clearly seen that some of the small micelles were aligned and
linked to strings, which obviously was the primary form of
nanoꢀfilament (nanoꢀfilament I). When the dialysis time was
increased to 12 h (Figure 4b), the nanoꢀfilaments could still be
observed, but more micelles were found attaching to the surꢀ
face of the nanoꢀfilaments, forming "branches" (nanoꢀfilament
II). In the same stage (Figure 4c), thin nanowires (nanowire I)
with rough surface and a wider diameter (ca. 10 nm) compared
to the primary nanoꢀfilaments were observed. These results
indicated that the nanoꢀfilaments were partially transformed
into nanowires via the growth of micelle "branch" on the filaꢀ
ment surface. Meanwhile, at this stage, single micelles or miꢀ
celle strings disappeared. Then from the sample taken at 24 h
(Figure 4d), wide and compact nanowires with a diameter
around 20~30 nm became the dominate morphology (nanꢀ
owire II). "Branched" structure was also found on the surface
of these nanowires, but here the "branch" turned to nanoꢀ
filaments with a diameter around 10 nm. These attached filaꢀ
ments were found twisted on the stem of the nanowire. Finalꢀ
ly, after 48 h (Figure 4e; Figure S10ꢀ11), these twisted nanoꢀ
filament "branches" became regularly circling the wire with a
period of 10 nm as shown in the inset of Figure 4e. Similar
regular structure was also observed in the assemblies of
Figure 3. The phase diagram of AAGP assemblies depends on
S/F ratio and different polypeptide scaffolds (P0, P1 and P2).
Although the architectures of our AAGB are complicated,
we inclined to think that the hydrophobicityꢀhydrophilicity
balance is still the main factor governing their behavior in selfꢀ
assembly. Therefore, we simply introduced a parameter S/F,
i.e. number ratio of saccharide units to phenylalanine monoꢀ
mer units in evaluating the assembly (Table 1). Thus a "phase
diagram" was constructed (Figure 3), in which S/F ratio and
the scaffolds character (P0, P1 and P2) serve the abscissa and
ordinate axes respectively. Fortunately, in the diagram, a clear
relationship between the assembly behavior and the structural
parameters of the library members displayed as follows. At
low S/F ratios (≤ 1.0), precipitates or irregular aggregates apꢀ
peared, probably because their relatively small hydrophilic
domains failed to stabilize the aggregates in water. When the
S/F ratio was increased to a much larger value (≥ 3.0), comꢀ
pound micelle became the major morphology, because of the
large hydrophilic and small hydrophobic domains of the polꢀ
ymers in this area. When S/F exists in the small window of 1.5
≤ S/F ≤ 3.0, nanowire was the only morphology observed. It
was noticed that at the upper boundary value 3.0 of S/F ratio,
P1tM-F under TEM (Figure S12) and of P2tM-F by cryꢀ
4
4
ofixation sample preparation method (Figure S13). At the
same time interval, thick and wide nanowires were also obꢀ
served (Figure 4f, nanowire III, diameter ca. 50 nm). Such
thick nanowires became more uniformity without inner strucꢀ
tures as probably a result of fusion of the surrounding nanoꢀ
filaments (Figure 4f; Figure S11).
stable nanowires were formed by P2tM-F with FFFF domain,
4
while compound micelles were formed by P1tM-F with FF
2
domain, so the longer phenylalanine had stronger regular
packing tendency leading to nanowires. In short, the selfꢀ
assembled morphologies were generally controlled by S/F
ratio, i.e. precipitates, nanoribbon, nanowires and compound
micelles were sequentially observed, when the S/F ratio was
increased.
Although the TEM studies could not trace the details of
the whole hierarchical selfꢀassembly process of AAGBs, the
evolution outline of the morphologies has been clearly
sketched. It becomes clear that the formation of the nanowires
of AAGBs stems from the basic assemblies i.e. the ordinary
micelles of AAGBs formed due to their amphiphilicity. The
following steps include the micelles alignment, formation of
the nanoꢀfilaments, attachment of micelles to the filaments,
formation of the nanowires due to the fusion of the attached
micelles and finally, formation of the thick nanowire etc. It is
not difficult to realize that the key factor driving all the steps is
the association between the sugar ‘shell’ of the primary miꢀ
Self-Assembly Mechanism of the AAGB Nanowires. The
nanowire, which was found in the small S/F "window", deꢀ
serves deep research because in previous selfꢀassembly studies
on glycopolypeptide, either peptide as main chain or pendent
groups, nanowire was rarely observed. However, in this work,
nanowires can be formed for all kinds of the main chain pepꢀ
tide, i.e. P0, P1 and P2 provided S/F exists in the window.
Therefore, we tried to use electronic microscopy to trace the
formation of the nanowires for understanding the mechanism.
42
celles, as well as the subsequent assemblies including the
nanoꢀfilaments, branches and nanowires. Such hierarchical
assembly has not been reported for other glycopolypeptides in
literature, probably because in our AAGBs, the coating hydroꢀ
philic layers of all of the assemblies at different levels consist
of much high sugar density due to the dendron structure of the
sugar pendants. The dense sugar shell might favor the
Nevertheless, formation of P1tM-F nanowires, as mentioned
4
above was quite fast (within a couple of hours), it was almost
not possible to capture stable intermediate states under TEM.
Considering that S/F of P2tM-F (3.0) was much higher than
4
that of P1tM-F (1.0), showing lower packing strength and
4
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Figure 4. TEM images of the nanowires formed by the hierarchical selfꢀassembly of P2tM-F and proposed selfꢀassembly mechanism, the
intermediate states captured from the samples prepared after (a) 3 h, (b, c) 12 h, (d) 24 h and (e, f) 48 h dialysis.
4
alignment of micelles forming string or nanoꢀfilament, which
became the second primary building block of the final nanꢀ
body 2G12 and plant lectin Concanavalin A (Con A) were
chosen as models to evaluate the protein binding ability of the
nanowires and compound micelles of AAGBs. 2G12 is a
broadly neutralizing human monoclonal antibody (mAb),
which can specifically recognize terminal Manα1ꢀ2Man moieꢀ
ties. First the binding ability of selfꢀassembled AAGBs to
2G12 was evaluated by sandwich Enzyme Linked Immunoꢀ
sorbent Assay (ELISA). In this experiment, 2G12 was immoꢀ
bilized to the surface coated with gp120, a major envelope
protein of HIV, which binds to 2G12 via the Nꢀlinked high
mannose glycan on its surface. Then inhibition of this bindꢀ
ing by adding free oligosaccharides and selfꢀassembled
AAGBs respectively was evaluated by ELISA. As shown in
Figure 5a, both Manα1ꢀ2Man (diMan), Manα1ꢀ2Manα1ꢀ2Man
(triMan) and their corresponding assemblies showed signifiꢀ
cant inhibition for 2G12ꢀcarbohydrate recognition, while
4
3
owire. In our previous study, we proposed that when two
micelles of block copolymers fused to each other, the chain
density of the shell on the two ends was lighter than that of the
lateral sides, which would direct the next coming micelle to
approach and attach to the two ends of the fused micelle. With
the heavy glycoꢀdendron shell in the current case, the alignꢀ
ment of micelles could be explained by the same rationale. On
the other hand, typical signal of βꢀsheet structure was observed
44
among the nanowire samples including P1tM-F , P2tM-F4
4
and P2dM-F by Circular dichroism (CD) spectroscopy (Figꢀ
4
ure S14), so βꢀsheet packing is another driving force leading to
nanowires. In contrast, CD spectra indicated that all compound
micelles exhibit typical random coil secondary structure rather
than βꢀsheet, and the nanowires formed by AAGBs with the
short peptide FF brushes show the same results (Figure S15ꢀ
mannose (Man) and its corresponding assembly P1M-F did
2
1
6). The result indicates that βꢀsheet forming tendency of oliꢀ
not. Furthermore, generally, compound micelles e.g. P2tM-F2
gophenylalanine is determined by the length of oligophenylalꢀ
anine species and is affected by the dense sugar shell.
showed higher inhibition ability to the binding between 2G12
and gp120 than nanowires e.g. P2tM-F did. These two polyꢀ
4
mer brushes shared the same generation of dendron, same
trisaccharide, but different morphologies and subsequently
different binding ability. The same phenomenon was also obꢀ
The effect of morphologies of AAGB assemblies of on their
binding ability to proteins. In nature, different morphologies
sometimes simply indicate different functions of the assemꢀ
blies. Herein, Human Immunodeficiency Virus (HIV) antiꢀ
served between P1tM-F
compound micelle and P1tM-F
2
4
nanowire. This may due to the less steric hindrance in comꢀ
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result indicated that by using a dense layer of oligosacchaꢀ
pound micelles than that in nanowires favoring the inhibition
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of the binding between 2G12 and oligosaccharides. Moreover,
when plant lectin Con A was utilized, this trend was even
more pronounced, as shown from the result of quantitative
agglutination experiment (Figure 5b), i.e. the compound
micelles gave a higher binding ability to Con A than nanꢀ
rides, new selfꢀassembly mechanism is possible, showing a
bright future of glycopolypeptide in mimicking nature and
developing new biocompatible materials. This newly syntheꢀ
sized AAGB architecture also brings us new thoughts in conꢀ
struction and the assembly mechanism of one dimensional
nanostructure.
4
5
owires did, especially between P2tM-F /P2tM-F and P1tM-
2
4
F /P1tM-F . These results might be explained by the different
2
4
curvature of the nanoꢀobjects, i.e. higher curvature of micelles
and lower ones of nanowires
ASSOCIATED CONTENT
Supporting Information
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Characterization of synthesized glycopolypeptide and correꢀ
1
sponding precursors, including HNMR and GPC results, dyꢀ
namic light scattering, circular dichroism, TEM, AFM, as well
as concentration dependent ELISA and UV agglutination reꢀ
sults are all available in supporting information.
AUTHOR INFORMATION
Corresponding Author guosong@fudan.edu.cn
ACKNOWLEDGMENT
National Natural Science Foundation of China (Nos.
9
1527305, 21474020, 91227203 and 51322306) and the Innoꢀ
vation Program of the Shanghai Municipal Education Comꢀ
mission are acknowledged for their financial support.
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67x196mm (150 x 150 DPI)
ACS Paragon Plus Environment

Page 13 of 17
Journal of the American Chemical Society
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50x206mm (150 x 150 DPI)
ACS Paragon Plus Environment

Journal of the American Chemical Society
Page 14 of 17
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209x209mm (150 x 150 DPI)
ACS Paragon Plus Environment

Page 15 of 17
Journal of the American Chemical Society
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336x262mm (150 x 150 DPI)
ACS Paragon Plus Environment

Journal of the American Chemical Society
Page 16 of 17
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39x230mm (150 x 150 DPI)
ACS Paragon Plus Environment

Page 17 of 17
Journal of the American Chemical Society
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07x334mm (150 x 150 DPI)
ACS Paragon Plus Environment