Tissue engineering, gene therapy, drug screening and emerging regenerative medicine therapies are fundamentally reliant on high-quality adherent cell culture, but current methods to cryopreserve cells in this format can give low cell yields and requires large volumes of solvent 'antifreezes'. Herein we report polyproline is a minimum (bio)synthetic mimic of antifreeze proteins, which is accessible by solution, solid phase and recombinant methods. We demonstrate that polyproline has ice recrystallization inhibition activity linked to its amphipathic helix and that it enhances the DMSO- cryopreservation of adherent cell lines. Polyproline may be a versatile additive in the emerging field of macromolecular cryoprotectants.
Multivalent Display of Antifreeze Proteins by Fusionto Self-Assembling Protein Cages Enhances Ice-Binding Activities
Multivalent Display of Antifreeze Proteins by Fusionto Self-Assembling Protein Cages Enhances Ice-Binding Activities
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/bichaw/0/bichaw.ahead-of-print/acs.biochem.6b00864/20161129/images/medium/bi-2016-00864x_0008.gif
Biochemistry
DOI: 10.1021/acs.biochem.6b00864
http://feeds.feedburner.com/~ff/acs/bichaw?d=yIl2AUoC8zA
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nmrlearner
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11-29-2016 09:02 PM
Comparison of backbone dynamics of the type III antifreeze protein and antifreeze-like domain of human sialic acid synthase
Comparison of backbone dynamics of the type III antifreeze protein and antifreeze-like domain of human sialic acid synthase
Abstract
Antifreeze proteins (AFPs) are found in a variety of cold-adapted (psychrophilic) organisms to promote survival at subzero temperatures by binding to ice crystals and decreasing the freezing temperature of body fluids. The type III AFPs are small globular proteins that consist of one α-helix, three 310-helices, and two β-strands. Sialic acids play important roles in a variety of biological functions, such as...
nmrlearner
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01-09-2015 03:58 PM
NMR characterizations of the ice binding surface of an antifreeze protein.
NMR characterizations of the ice binding surface of an antifreeze protein.
NMR characterizations of the ice binding surface of an antifreeze protein.
PLoS One. 2010;5(12):e15682
Authors: Hong J, Hu Y, Li C, Jia Z, Xia B, Jin C
Antifreeze protein (AFP) has a unique function of reducing solution freezing temperature to protect organisms from ice damage. However, its functional mechanism is not well understood. An intriguing question concerning AFP function is how the high selectivity for ice ligand is achieved in the presence of free water of...
nmrlearner
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01-07-2011 11:21 PM
[NMR paper] Protein structure elucidation from minimal NMR data: the CLOUDS approach.
Protein structure elucidation from minimal NMR data: the CLOUDS approach.
Related Articles Protein structure elucidation from minimal NMR data: the CLOUDS approach.
Methods Enzymol. 2005;394:261-95
Authors: Grishaev A, Llinás M
In this chapter we review automated methods of protein NMR data analysis and expand on the assignment-independent CLOUDS approach. As presented, given a set of reliable NOEs it is feasible to derive a spatial H-atom distribution that provides a low-resolution image of the protein structure. In order to generate such a...
nmrlearner
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11-24-2010 11:14 PM
[NMR paper] Binding of copper(II) ions to the polyproline II helices of PEVK modules of the giant
Binding of copper(II) ions to the polyproline II helices of PEVK modules of the giant elastic protein titin as revealed by ESI-MS, CD, and NMR.
Related Articles Binding of copper(II) ions to the polyproline II helices of PEVK modules of the giant elastic protein titin as revealed by ESI-MS, CD, and NMR.
Biopolymers. 2003 Oct;70(3):297-309
Authors: Ma K, Wang K
Titin, a family of giant elastic proteins, constitutes an elastic sarcomere matrix in striated muscle. In the I-band region of the sarcomere, the titin PEVK segment acts as a molecular...
nmrlearner
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11-24-2010 09:16 PM
[NMR paper] Automated protein fold determination using a minimal NMR constraint strategy.
Automated protein fold determination using a minimal NMR constraint strategy.
Related Articles Automated protein fold determination using a minimal NMR constraint strategy.
Protein Sci. 2003 Jun;12(6):1232-46
Authors: Zheng D, Huang YJ, Moseley HN, Xiao R, Aramini J, Swapna GV, Montelione GT
Determination of precise and accurate protein structures by NMR generally requires weeks or even months to acquire and interpret all the necessary NMR data. However, even medium-accuracy fold information can often provide key clues about protein evolution...
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11-24-2010 09:01 PM
Protein-ice interaction of an antifreeze protein observed with solid-state NMR [Chemi
Protein-ice interaction of an antifreeze protein observed with solid-state NMR
Siemer, A. B., Huang, K.-Y., McDermott, A. E....
Date: 2010-10-12
NMR on frozen solutions is an ideal method to study fundamental questions of macromolecular hydration, because the hydration shell of many biomolecules does not freeze together with bulk solvent. In the present study, we present previously undescribed NMR methods to study the interactions of proteins with their hydration shell and the ice lattice in frozen solution. We applied these methods to compare solvent interaction of an ice-binding...
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10-13-2010 04:10 AM
Protein-ice interaction of an antifreeze protein observed with solid-state NMR.
Protein-ice interaction of an antifreeze protein observed with solid-state NMR.
Related Articles Protein-ice interaction of an antifreeze protein observed with solid-state NMR.
Proc Natl Acad Sci U S A. 2010 Sep 30;
Authors: Siemer AB, Huang KY, McDermott AE
NMR on frozen solutions is an ideal method to study fundamental questions of macromolecular hydration, because the hydration shell of many biomolecules does not freeze together with bulk solvent. In the present study, we present previously undescribed NMR methods to study the interactions...