Dynamic protein–water relationships during β-sheet formation
Abstract
We investigated the polymer−water interaction in a model fibrous protein. Bombyx mori silk fibroin film, a typical model of biodegradable material, was cast from aqueous solution and analyzed in this study. Differential scanning calorimetry (DSC), its temperature-modulated variant (TMDSC), and the time-resolved technique of Fourier transform infrared spectroscopy (FTIR) were used for the first time to monitor the detailed structural changes of silk fibroin during heating and during isothermal crystallization above the glass transition temperature, T g ∼ 451 K (178 °C). Results show that intermolecular bound water molecules, acting as a plasticizer, will strongly affect the secondary structure of silk fibroin. DSC study shows that silk fibroin initially displays a water-induced glass transition around 80 °C during heating, resulting from a temporary bound water-silk structure. Quantitative thermal analysis of the heat capacity changes of this system during heating revealed that no β-sheet crystals were formed below T g . FTIR scans also confirmed that no β-sheets were formed during heating and removal of bound water below T g . During water removal, the amide II region shifts to lower frequency, and the 1515 cm −1 band increases slightly, indicating change of the microenvironment in the silk fibroin chains. During isothermal crystallization above T g, the amide I region of silk fibroin spectra showed a phase transition from the secondary structures of noncrystalline random coils and α-helices to the β-pleated-sheet crystals, while the amide II region and the 1515 cm −1 band remain stable during β-sheet formation. Analogy is made between the crystallization of synthetic polymers according to the four-state scheme of Strobl and the crystallization process of silk fibroin, which we suggest proceeds through an intermediate precursor stage associated with water removal. This study provides a deeper understanding of the formation of β-pleated sheets during the crystallization process in silk fibroin, with implications for the crystallization of naturally occurring silk fibers from animals such as the silkworm and spider.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
A low-temperature endothermic event in SFMA at approximately 90.0 °C is attributed to relaxation phenomena involving bound water and chemically modified chain segments.
“A low-temperature endothermic event observed at approximately 90.0 °C is attributed to relaxation phenomena involving bound water and chemically modified chain segments [48].”
Cites
Altman, G. H., Diaz, F., Jakuba, C., Calabro, T., Horan, R. L., Chen, J., Lu, H., Richmond, J., & Kaplan, D. L. (2003) · Biomaterials · doi:10.1016/s0142-9612(02)00353-8 · Cited by 3,458 · see citing works
Self-citationWhat vibrations tell about proteins
Barth, A., & Zscherp, C. (2002) · Quarterly Reviews of Biophysics · doi:10.1017/s0033583502003815 · Cited by 2,088 · see citing works
Mechanism of silk processing in insects and spiders
Jin, H., & Kaplan, D. L. (2003) · Nature · doi:10.1038/nature01809 · Cited by 1,379 · see citing works
Self-citationThe infrared absorption of amino acid side chains
Barth, A. (2000) · Progress in Biophysics and Molecular Biology · doi:10.1016/s0079-6107(00)00021-3 · Cited by 1,353 · see citing works
Protein secondary structures in water from second-derivative amide I infrared spectra
Dong, A., Huang, P., & Caughey, W. S. (1990) · Biochemistry · doi:10.1021/bi00465a022 · Cited by 1,333 · see citing works
Hu, X., Kaplan, D. L., & Cebe, P. (2006) · Macromolecules · doi:10.1021/ma0610109 · Cited by 1,251 · see citing works
Self-citationHydrogen bonding in polymers: infrared temperature studies of an amorphous polyamide
Skrovanek, D. J., Howe, S. E., Painter, P. C., et al. (1985) · Macromolecules · doi:10.1021/ma00151a006 · Cited by 623 · see citing works
Venyaminov, S. Y., & Kalnin, N. N. (1990) · Biopolymers · doi:10.1002/bip.360301309 · Cited by 590 · see citing works
Goormaghtigh, E., Cabiaux, V., & Ruysschaert, J. (1990) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1990.tb19354.x · Cited by 538 · see citing works
Strobl, G. R. (2000) · The European Physical Journal E · doi:10.1007/s101890070030 · Cited by 520 · see citing works
Murayama, K., & Tomida, M. (2004) · Biochemistry · doi:10.1021/bi0489154 · Cited by 482 · see citing works
Wunderlich, B., Jin, Y., & Boller, A. (1994) · Thermochimica Acta · doi:10.1016/s0040-6031(94)85214-6 · Cited by 444 · see citing works
Structural characteristics and properties of the regenerated silk fibroin prepared from formic acid
Um, I. C., Kweon, H., Park, Y. H., & Hudson, S. M. (2001) · International Journal of Biological Macromolecules · doi:10.1016/s0141-8130(01)00159-3 · Cited by 436 · see citing works
Calorimetric Study of the Glassy State. IV. Heat Capacities of Glassy Water and Cubic Ice
Sugisaki, M., Suga, H., & Seki, S. (1968) · Bulletin of the Chemical Society of Japan · doi:10.1246/bcsj.41.2591 · Cited by 431 · see citing works
Hydrogen bonding in polymers. 2. Infrared temperature studies of nylon 11
Skrovanek, D. J., Painter, P. C., & Coleman, M. M. (1986) · Macromolecules · doi:10.1021/ma00157a037 · Cited by 423 · see citing works
Kim, Y. S., Dong, L., Hickner, M. A., Glass, T. E., Webb, V., & McGrath, J. E. E. (2003) · Macromolecules · doi:10.1021/ma0301451 · Cited by 387 · see citing works
Cai, S., & Singh, B. R. (1999) · Biophysical Chemistry · doi:10.1016/s0301-4622(99)00060-5 · Cited by 360 · see citing works
Free Volume and the Mechanism of Plasticization in Water-Swollen Poly(vinyl alcohol)
Hodge, R. M., Bastow, T. J., Edward, G. H., Simon, G. P., & Hill, A. J. (1996) · Macromolecules · doi:10.1021/ma951073j · Cited by 349 · see citing works
Cai, S., & Singh, B. R. (2004) · Biochemistry · doi:10.1021/bi030149y · Cited by 313 · see citing works
Venyaminov, S. Y., & Kalnin, N. N. (1990) · Biopolymers · doi:10.1002/bip.360301310 · Cited by 294 · see citing works
Chen, X., Shao, Z., Marinković, N., et al. (2001) · Biophysical Chemistry · doi:10.1016/s0301-4622(00)00213-1 · Cited by 293 · see citing works
Jung, C. (2000) · Journal of Molecular Recognition · doi:10.1002/1099-1352(200011/12)13:6 · Cited by 245 · see citing works
Genetic engineering of fibrous proteins: spider dragline silk and collagen
Foo, C. W. P., & Kaplan, D. L. (2002) · Advanced Drug Delivery Reviews · doi:10.1016/s0169-409x(02)00061-3 · Cited by 208 · see citing works
Self-citationEffect of water on the thermal properties of silk fibroin
Hu, X., Kaplan, D. L., & Cebe, P. (2006) · Thermochimica Acta · doi:10.1016/j.tca.2006.12.011 · Cited by 202 · see citing works
Self-citationFourier transform Raman and Fourier transform infrared spectroscopy studies of silk fibroin
Shao, J., Zheng, J., Liu, J., et al. (2005) · Journal of Applied Polymer Science · doi:10.1002/app.21346 · Cited by 198 · see citing works
Protein–water displacement distributions
Doster, W., & Settles, M. (2005) · Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics · doi:10.1016/j.bbapap.2005.03.010 · Cited by 194 · see citing works
Fu, F.-N., DeOliveira, D. B., Trumble, W. R., Sarkar, H. K., & Singh, B. R. (1994) · Applied Spectroscopy · doi:10.1366/0003702944028065 · Cited by 191 · see citing works
Ishida, M., Asakura, T., Yokoi, M., et al. (1990) · Macromolecules · doi:10.1021/ma00203a017 · Cited by 191 · see citing works
Mikhonin, A. V., Bykov, S. V., Myshakina, N. S., & Asher, S. A. (2006) · The Journal of Physical Chemistry B · doi:10.1021/jp054593h · Cited by 174 · see citing works
The role of formic acid in solution stability and crystallization of silk protein polymer
Um, I. C., Kweon, H., Lee, K. G., et al. (2003) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2003.08.004 · Cited by 170 · see citing works
Tretinnikov, O. N., & Tamada, Y. (2001) · Langmuir · doi:10.1021/la010791y · Cited by 170 · see citing works
van Stokkum, I. H. M., Linsdell, H., Hadden, J. M., et al. (1995) · Biochemistry · doi:10.1021/bi00033a024 · Cited by 163 · see citing works
Um, I. C., Ki, C. S., Kweon, H., et al. (2004) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2004.03.011 · Cited by 159 · see citing works
Raman spectroscopic studies of silk fibroin fromBombyx mori
Monti, P., Freddi, G., Bertoluzza, A., et al. (1998) · Journal of Raman Spectroscopy · doi:10.1002/(sici)1097-4555(199804)29:4 · Cited by 154 · see citing works
Fabian, H., Schultz, C., Naumann, D., et al. (1993) · Journal of Molecular Biology · doi:10.1006/jmbi.1993.1442 · Cited by 153 · see citing works
Min, B., Jeong, L., Lee, K. Y., & Park, W. H. (2006) · Macromolecular Bioscience · doi:10.1002/mabi.200500246 · Cited by 151 · see citing works
Effect of moisture absorption on the thermal properties ofBombyx mori silk fibroin films
Agarwal, N., Hoagland, D. A., & Farris, R. J. (1997) · Journal of Applied Polymer Science · doi:10.1002/(sici)1097-4628(19970118)63:3 · Cited by 149 · see citing works
Structural Study of Irregular Amino Acid Sequences in the Heavy Chain of Bombyx m ori Silk Fibroin
Ha, S.-W., Gracz, H. S., Tonelli, A. E., & Hudson, S. M. (2005) · Biomacromolecules · doi:10.1021/bm050294m · Cited by 142 · see citing works
Xu, H., & Cebe, P. (2004) · Macromolecules · doi:10.1021/ma035961n · Cited by 139 · see citing works
Self-citationThermally Induced α-Helix to β-Sheet Transition in Regenerated Silk Fibers and Films
Drummy, L. F., Phillips, D. M., Stone, M. O., et al. (2005) · Biomacromolecules · doi:10.1021/bm0503524 · Cited by 138 · see citing works
McGrath, K. P., & Kaplan, D. L. (1997) · Birkhäuser Boston eBooks · doi:10.1007/978-1-4612-4094-5 · Cited by 128 · see citing works
Self-citationCrystallinity in silk fibers: Partial acid hydrolysis and related studies
Bhat, N. V., & Nadiger, G. S. (1980) · Journal of Applied Polymer Science · doi:10.1002/app.1980.070250518 · Cited by 122 · see citing works
Schmidtke, J., Strobl, G. R., & Thurn‐Albrecht, T. (1997) · Macromolecules · doi:10.1021/ma9703923 · Cited by 117 · see citing works
Magoshi, J., Magoshi, Y., Nakamura, S., et al. (1977) · Journal of Polymer Science Polymer Physics Edition · doi:10.1002/pol.1977.180150915 · Cited by 117 · see citing works
Methods to study protein folding by stopped-flow FT-IR
Fabian, H. (2004) · Methods · doi:10.1016/j.ymeth.2004.03.004 · Cited by 109 · see citing works
Hienerwadel, R., Boussac, A., Breton, J., Diner, B. A., & Berthomieu, C. (1997) · Biochemistry · doi:10.1021/bi971521a · Cited by 108 · see citing works
Slade, L., & Levine, H. (1995) · Journal of Food Engineering · doi:10.1016/0260-8774(95)90766-5 · Cited by 104 · see citing works
Jeong, L., Lee, K. Y., Liu, J. W., et al. (2006) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2006.02.009 · Cited by 101 · see citing works
Reinstädler, D., Fabian, H., Backmann, J., & Naumann, D. (1996) · Biochemistry · doi:10.1021/bi961810j · Cited by 98 · see citing works
Kinetic modeling of the chain-end functionality in atom transfer radical polymerization
Lutz, J., & Matyjaszewski, K. A. (2002) · Macromolecular Chemistry and Physics · doi:10.1002/1521-3935(200207)203:10/11 · Cited by 91 · see citing works
Fabian, H., Schultz, C., Backmann, J., Hahn, U., Saenger, W., Mantsch, H. H., & Naumann, D. (1994) · Biochemistry · doi:10.1021/bi00201a021 · Cited by 91 · see citing works
DSC studies on bound water in silk fibroin/S-carboxymethyl kerateine blend films
Lee, K. Y., & Ha, W. (1999) · Polymer · doi:10.1016/s0032-3861(98)00611-9 · Cited by 77 · see citing works
Methionine Redox Controlled Crystallization of Biosynthetic Silk Spidroin
Valluzzi, R., Szela, S., Avtges, P., et al. (1999) · The Journal of Physical Chemistry B · doi:10.1021/jp991363s · Cited by 71 · see citing works
Computation of Heat Capacities of Liquid Polymers
Pyda, M., & Wunderlich, B. (1999) · Macromolecules · doi:10.1021/ma9816620 · Cited by 68 · see citing works
Backbone dynamics, fast folding, and secondary structure formation in helical proteins and peptides
Hardin, C., Luthey-Schulten, Z. A., & Wolynes, P. G. (1999) · Proteins Structure Function and Bioinformatics · doi:10.1002/(sici)1097-0134(19990215)34:3 · Cited by 59 · see citing works
Direct UV Raman Monitoring of 3 10 -Helix and π-Bulge Premelting during α-Helix Unfolding
Mikhonin, A. V., & Asher, S. A. (2006) · Journal of the American Chemical Society · doi:10.1021/ja062269+ · Cited by 53 · see citing works
Conformational contribution to the heat capacity of the starch and water system
Pyda, M. (2001) · Journal of Polymer Science Part B Polymer Physics · doi:10.1002/polb.10060 · Cited by 52 · see citing works
Yamane, T., Umemura, K., Nakazawa, Y., et al. (2003) · Macromolecules · doi:10.1021/ma034466o · Cited by 52 · see citing works
Conformational Heat Capacity of Interacting Systems of Polymer and Water
Pyda, M. (2002) · Macromolecules · doi:10.1021/ma0118466 · Cited by 46 · see citing works
Lin, S., & Chu, H.-L. (2003) · International Journal of Biological Macromolecules · doi:10.1016/s0141-8130(03)00051-5 · Cited by 43 · see citing works
Wang, S., Lin, S., Li, M., et al. (2004) · Biophysical Chemistry · doi:10.1016/j.bpc.2004.12.004 · Cited by 39 · see citing works
Pearce, R., & Vancso, G. J. (1998) · Journal of Polymer Science Part B Polymer Physics · doi:10.1002/(sici)1099-0488(199810)36:14 · Cited by 38 · see citing works
Crystalline Regions of Bombyx mori Silk Fibroin May Exhibit β-Turn and β-Helix Conformations
Lazo, N. D., & Downing, D. T. (1999) · Macromolecules · doi:10.1021/ma9900582 · Cited by 37 · see citing works
Sonoyama, M., & Nakano, T. (2000) · Applied Spectroscopy · doi:10.1366/0003702001950698 · Cited by 36 · see citing works
Cell asymmetry correction for temperature modulated differential scanning calorimetry
Ishikiriyama, K., & Wunderlich, B. (1997) · Journal of thermal analysis · doi:10.1007/bf01980494 · Cited by 34 · see citing works
Determination of cell asymmetry in temperature-modulated DSC
Boller, A., Okazaki, I., Ishikiriyama, K., et al. (1997) · Journal of thermal analysis · doi:10.1007/bf01996796 · Cited by 31 · see citing works
X-ray Evidence for a “Super”-Secondary Structure in Silk Fibers
Valluzzi, R., & Jin, H. (2004) · Biomacromolecules · doi:10.1021/bm0343085 · Cited by 30 · see citing works
Michaux, M.-A., Dautant, A., Gallois, B., Granier, T., d’Estaintot, B. L., & PRECIGOUX, G. (1996) · Proteins Structure Function and Bioinformatics · doi:10.1002/(sici)1097-0134(199603)24:3 · Cited by 21 · see citing works
Stellwagen, J., & Stellwagen, N. C. (1994) · Biopolymers · doi:10.1002/bip.360340914 · Cited by 16 · see citing works
Heat Capacity Study of Solution Grown Crystals of Isotactic Polystyrene
Xu, H., & Cebe, P. (2005) · Macromolecules · doi:10.1021/ma048042l · Cited by 15 · see citing works
Self-citationLin, S., Hsieh, T., & Wei, Y. (2005) · Peptides · doi:10.1016/j.peptides.2004.11.005 · Cited by 11 · see citing works
Simple solution approach to hollow nanospheres of cubic HgS
Jian-quan, L., Feng, Y., Zhang, S., et al. (2005) · Materials Letters · doi:10.1016/j.matlet.2005.05.028 · Cited by 7 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Water-insoluble silk films with silk I structure
Lu, Q., Hu, X., Wang, X., Kluge, J. A., Lu, S., Cebe, P., & Kaplan, D. L. (2010) · Acta Biomaterialia · doi:10.1016/j.actbio.2009.10.041 · Cited by 654 · see citing works
Self-citationDegradation mechanism and control of silk fibroin
Lu, Q., Zhang, B., Li, M., Zuo, B., Kaplan, D. L., Huang, Y., & Zhu, H. (2011) · Biomacromolecules · doi:10.1021/bm101422j · Cited by 344 · see citing works
Self-citationAntunes, B. S. L., Pedrini, F., Komatsu, D., Asami, J., Dicko, C., & Duek, E. A. R. (2026) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2026.154129
- Authors
- Hu, X., Kaplan, D., & Cebe, P.
- Affiliations
- Tufts University (US)
- Corresponding author
- Cebe, Peggy
- Venue
- Macromolecules
- Publisher
- American Chemical Society (ACS)
- Published
- 2008-05-10
- Volume
- 41
- Issue
- 11
- Pages
- 3939-3948
- ISSN
- 0024-9297 (ISSN-L)
- 1520-5835
- OpenAlex
- W2002787011
- Cited by
- 315
- References
- 72
- Citations by year
- 2026: 18 · 2025: 22 · 2024: 15 · 2023: 19 · 2022: 17 · 2021: 20 · 2020: 12 · 2019: 19 · 2018: 17 · 2017: 29 · 2016: 17 · 2015: 12 · 2014: 25 · 2013: 15 · 2012: 17
- FWCI
- 5.89
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Macromolecules (publishedVersion)
- Topics
- Physical Sciences › Materials Science › Biomaterials › Silk-based biomaterials and applications
- Physical Sciences › Materials Science › Polymers and Plastics › Polymer crystallization and properties
- Physical Sciences › Materials Science › Materials Chemistry › Material Dynamics and Properties
- Keywords
- silk fibroin, β-sheet formation, polymer-water interactions, bound water, plasticizer effect, glass transition, isothermal crystallization, heat capacity, alpha-helix, protein secondary structure, Bombyx mori
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
