Dynamic Stabilization of Expressed Proteins in Engineered Diatom Biosilica Matrices Public Deposited

http://ir.library.oregonstate.edu/concern/articles/tt44pp34v

To the best of our knowledge, one or more authors of this paper were federal employees when contributing to this work. This is the publisher’s final pdf. The published article is copyrighted by the American Chemical Society and can be found at:  http://pubs.acs.org/journal/bcches

Descriptions

Attribute NameValues
Creator
Abstract or Summary
  • Self-assembly of recombinant proteins within the biosilica of living diatoms represents a means to construct functional materials in a reproducible and scalable manner that will enable applications that harness the inherent specificities of proteins to sense and respond to environmental cues. Here we describe the use of a silaffin-derived lysine-rich 39-amino-acid targeting sequence (Sil3[subscript]T8) that directs a single chain fragment variable (scFv) antibody or an enhanced green fluorescent protein (EGFP) to assemble within the biosilica frustule, resulting in abundance of >200 000 proteins per frustule. Using either a fluorescent ligand bound to the scFv or the intrinsic fluorescence of EGFP, we monitored protein conformational dynamics, accessibility to external quenchers, binding affinity, and conformational stability. Like proteins in solution, proteins within isolated frustules undergo isotropic rotational motion, but with 2-fold increases in rotational correlation times that are indicative of weak macromolecular associations within the biosilica. Solvent accessibilities and high-affinity (pM) binding are comparable to those in solution. In contrast to solution conditions, scFv antibodies within the biosilica matrix retain their binding affinity in the presence of chaotropic agents (i.e., 8 M urea). Together, these results argue that dramatic increases in protein conformational stability within the biosilica matrices arise through molecular crowding, acting to retain native protein folds and associated functionality with the potential to allow the utility of engineered proteins under a range of harsh environmental conditions associated with environmental sensing and industrial catalytic transformations.
Resource Type
DOI
Date Available
Date Issued
Citation
  • Xiong, Y., Ford, N. R., Hecht, K. A., Roesijadi, G., & Squier, T. C. (2016). Dynamic Stabilization of Expressed Proteins in Engineered Diatom Biosilica Matrices. Bioconjugate Chemistry, 27(5), 1205-1209. doi:10.1021/acs.bioconjchem.6b00165
Series
Rights Statement
Funding Statement (additional comments about funding)
Publisher
Peer Reviewed
Language
Replaces
Additional Information
  • description.provenance : Submitted by Patricia Black (patricia.black@oregonstate.edu) on 2016-06-30T20:51:50Z No. of bitstreams: 2 XiongDynamicStabilizationExpressed.pdf: 697553 bytes, checksum: ad03ec09074ce0fd18a79ea3713c336a (MD5) XiongDynamicStabilizationExpressedSupportingInformation.pdf: 576038 bytes, checksum: 998ed96de1c374cc1b9ac98178f0c7a2 (MD5)
  • description.provenance : Made available in DSpace on 2016-06-30T20:52:09Z (GMT). No. of bitstreams: 2 XiongDynamicStabilizationExpressed.pdf: 697553 bytes, checksum: ad03ec09074ce0fd18a79ea3713c336a (MD5) XiongDynamicStabilizationExpressedSupportingInformation.pdf: 576038 bytes, checksum: 998ed96de1c374cc1b9ac98178f0c7a2 (MD5) Previous issue date: 2016-05
  • description.provenance : Approved for entry into archive by Patricia Black(patricia.black@oregonstate.edu) on 2016-06-30T20:52:09Z (GMT) No. of bitstreams: 2 XiongDynamicStabilizationExpressed.pdf: 697553 bytes, checksum: ad03ec09074ce0fd18a79ea3713c336a (MD5) XiongDynamicStabilizationExpressedSupportingInformation.pdf: 576038 bytes, checksum: 998ed96de1c374cc1b9ac98178f0c7a2 (MD5)

Relationships

In Administrative Set:
Last modified: 07/26/2017 Default
Citations:

EndNote | Zotero | Mendeley

Items