harnessing peptide binding to capture and reclaim phosphate
TRANSCRIPT
doi.org/10.26434/chemrxiv.12771623.v1
Harnessing Peptide Binding to Capture and Reclaim PhosphateWhitney Fowler, Chuting Deng, Gabriella Griffen, Tess Teodoro, Ashley Z. Guo, Michal Zaiden, MosheGottlieb, Juan de Pablo, Matthew Tirrell
Submitted date: 06/08/2020 • Posted date: 07/08/2020Licence: CC BY-NC-ND 4.0Citation information: Fowler, Whitney; Deng, Chuting; Griffen, Gabriella; Teodoro, Tess; Guo, Ashley Z.;Zaiden, Michal; et al. (2020): Harnessing Peptide Binding to Capture and Reclaim Phosphate. ChemRxiv.Preprint. https://doi.org/10.26434/chemrxiv.12771623.v1
With rising consumer demands, society is tapping into wastewater as an innovative source to recycledepleting resources. Novel reclamation technologies have been recently explored for this purpose, includingseveral that optimize natural biological processes for targeted reclamation. However, this emerging field has anoticeable dearth of synthetic material technologies that are programmed to capture, release and recyclespecified targets, and of the novel materials that do exist, synthetic platforms incorporating biologicallyinspired mechanisms are rare. We present here a prototype of a materials platform utilizing peptideamphiphiles that has been molecularly engineered to sequester, release, and reclaim phosphate utilizing astimuli-responsive pH trigger, exploiting a protein-inspired binding mechanism that is incorporated directly intothe self-assembled material network. This material is able to sequester completely and controllably releasephosphate for multiple cycles of reuse. We have determined by simulations that the binding conformation ofthe peptide becomes constrained in the dense micelle corona at high pH such that phosphate is expelledwhen it otherwise would be preferentially bound. However, at neutral pH, this dense structure converselyemploys multi-chain binding to further stabilize phosphate when it would otherwise be unbound, openingopportunities for higher-order conformational binding design to be engineered into this controllably packedcorona. With this work, we are pioneering a new platform to be readily altered to capture other valuabletargets, presenting a new class of capture and release materials for recycling resources on the nanoscale.
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2 3 4 5 6 7 8 9 10 110%
50%
100%
PO
4 B
ound (
%)
pH
A
2 3 4 5 6 7 8 9 10 11pH
B
1:1 2:1 3:1 5:10%
50%
100%
PO
4 B
ound (
%)
PA Binding Unit: PO4
C
1:1 5:1 10:1
PA Binding Unit: PO4
D
C16GSHhexC16GGGhex
ASSOCIATED CONTENT
Supporting Information
AUTHOR INFORMATION
Corresponding Authors
ACKNOWLEDGMENT
REFERENCES
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S-1
Supporting Information
Harnessing Peptide Binding to Capture and Reclaim Phosphate
Table of Contents
S-2
1. Additional Experimental Results and Discussion
1.1 Mass Spectroscopy Verification using Matrix-Assisted Laser Desorption-Ionization – Time of
Flight (MALDI-TOF)
1.2 Purity Analysis using Liquid Chromatography – Mass Spectroscopy (LC-MS)
S-7
1.5 Spectrophotometric Molybdenum Blue Assay
0 10 20 30 40 500.0
0.1
0.2
0.3
0.4
0.5
0.6
Ab
sorb
an
ce
(a
.u.)
Concentration of Phosphate (ppm)
R2 = 0.999
S-8
1.6 Analysis of Kinetics of Binding
1.7 Effect of Increased NaCl on Binding and the Molybdenum Blue Assay
0 1 2
0
25
50[P
O4]
Bo
un
d (
pp
m)
Time (h)
pH 6
pH 2
pH 11
Avg. = 46.64 ppm
Avg. = 5.35 ppm
Avg. = 2.45 ppm 0%
25%
50%
75%
100%
PO
4 B
ou
nd
(%
)
S-10
2. Additional Simulation Results and Discussion
2.1 Estimated Potential of Mean Force for Phosphate Micelle Binding
PMF(r) =
kTln(P(r))
2.2 Representative Snapshot of Multi-chain Binding
S-11
3. Method and Materials
3.1 Synthesis and Purification of Peptide Amphiphile Micelles and Micelle Preparation Procedure
S-12
3.3 Negative-stain Transmission Electron Microscopy (TEM) Imaging
3.4 Molybdenum Blue Assay for Analyzing Phosphate in Solution
3.5 Analysis of pH-Dependent Phosphate Binding
S-14
3.8 Analysis of the Effect of NaCl on Binding and the Molybdenum Blue Assay
3.9 Simulation Model and Force Field Parameters
𝑖 𝑗
𝑢𝑛𝑏(𝑟𝑖𝑗) = 4𝜖𝑖𝑗 [(𝜎𝑖𝑗
𝑟𝑖𝑗)
12
− (𝜎𝑖𝑗
𝑟𝑖𝑗)
6
] +𝑞𝑖𝑞𝑗
4𝜋𝜀0𝑟𝑖𝑗,
𝑟𝑖𝑗 𝑖 𝑗 𝜎𝑖𝑗 𝜎𝑖𝑗
𝜀0 𝑞𝑖 𝑞𝑗 𝑖 𝑗
S-15
𝑚 𝜎𝑖𝑖 nm 𝜖𝑖𝑖kJ
mol𝑞 (𝑒)
𝑢𝑏𝑜𝑛𝑑(𝑟𝑖𝑗) =𝑘𝑏𝑜𝑛𝑑
2(𝑟𝑖𝑗 − 𝑟𝑖𝑗
(0))2
,
𝑟𝑖𝑗 𝑖 𝑗 𝑘𝑏𝑜𝑛𝑑 𝑟𝑖𝑗(0)
𝑟𝑖𝑗(0)(nm) 𝑘𝑏𝑜𝑛𝑑
kJ
mol∙nm2
𝑢𝑏𝑒𝑛𝑑(𝜃𝑖𝑗𝑘) = 𝑘𝑏𝑒𝑛𝑑(𝜃𝑖𝑗𝑘 − 𝜃𝑖𝑗𝑘(0))
2,
𝑘𝑏𝑒𝑛𝑑 𝜃𝑖𝑗𝑘 𝑖 𝑗 𝑘 𝜃𝑖𝑗𝑘(0)
S-16
𝜃𝑖𝑗𝑘(0)(deg) 𝑘𝑏𝑒𝑛𝑑
kJ
mol∙rad2
The torsional potentials for 1-2-3-4 bonded united atoms take the form:
𝑢𝑡𝑜𝑟𝑠(𝜙𝑖𝑗𝑘𝑙) = 𝑘𝜙[1 + cos(𝑛 𝜙𝑖𝑗𝑘𝑙 − 𝜙𝑖𝑗𝑘𝑙(0))] (4)
where 𝑘𝜙 is the coefficient and 𝜙𝑖𝑗𝑘𝑙 is the dihedral angle defined by atoms 𝑖, 𝑗, 𝑘, and 𝑙. 𝑛 is the integer
multiplicity and 𝜙𝑖𝑗𝑘𝑙(0)
is a refence dihedral angle. The torsional interaction parameters are listed in Table S7.
𝜙𝑖𝑗𝑘𝑙(0)(deg) 𝑘𝜙
kJ
mol𝑛
3.10 Simulations of Phosphate Binding to Single-Chain
S-17
3.11 Simulations of Phosphate Binding to Peptide Amphiphile Micelle
3.12 Safety Comment
4. Author Roles and Responsibilities
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