Cell-free expression and characterization of multivalent rhamnose-binding lectins using bio-layer interferometry.

Publisher: IRL Press at Oxford University Press Country of Publication: England NLM ID: 9104124 Publication Model: Print Cited Medium: Internet ISSN: 1460-2423 (Electronic) Linking ISSN: 09596658 NLM ISO Abbreviation: Glycobiology Subsets: MEDLINE

Original Publication: Oxford ; New York : IRL Press at Oxford University Press, c1990-

Lectins*/chemistry ; Rhamnose*; Carbohydrates/chemistry ; Recombinant Proteins/genetics ; Interferometry/methods

Lectins are important biological tools for binding glycans, but recombinant protein expression poses challenges for some lectin classes, limiting the pace of discovery and characterization. To discover and engineer lectins with new functions, workflows amenable to rapid expression and subsequent characterization are needed. Here, we present bacterial cell-free expression as a means for efficient, small-scale expression of multivalent, disulfide bond-rich, rhamnose-binding lectins. Furthermore, we demonstrate that the cell-free expressed lectins can be directly coupled with bio-layer interferometry analysis, either in solution or immobilized on the sensor, to measure interaction with carbohydrate ligands without purification. This workflow enables the determination of lectin substrate specificity and estimation of binding affinity. Overall, we believe that this method will enable high-throughput expression, screening, and characterization of new and engineered multivalent lectins for applications in synthetic glycobiology.
(© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Keywords: bio-layer interferometry; cell-free production; lectin

0 (Lectins)
QN34XC755A (Rhamnose)
0 (Carbohydrates)
0 (Recombinant Proteins)

Date Created: 20230307 Date Completed: 20230608 Latest Revision: 20230608

20240628

10.1093/glycob/cwad018

36882003