Study and characterization of the interactions of the recently discovered functional amyloid fibril, FapC with signaling molecules and potential inhibitors with three biosensors : Biacore™ T200, EnSpire® and LigandTracer®.

• Main Author: Gea, Chong Jin.
• Other Authors: Susana Geifman Shochat
• Format: Final Year Project
• Language: English
• Published: 2013
• Subjects: DRNTU::Science::Biological sciences::Microbiology::Bacteria; DRNTU::Science::Biological sciences::Biophysics
• Online Access: http://hdl.handle.net/10356/52908
• Institution: Nanyang Technological University

Pseudomonas spp. biofilms typically grow in aqueous environments of high shear and flow rate. Despite the conditions, the biofilms are able to retain signaling molecules that are involved in QS and biofilm development through EPS. While it is unknown how these molecules bind, it has been proposed that these signaling molecules are retained by an amyloid fibril, FapC, found in the EPS. The Biacore™ T200, EnSpire® and LigandTracer® were employed to study and characterize the interactions of these signaling molecules and also of potential inhibitors of biofilm formation with FapC. The results from the T200 indicated that the signaling molecules bound to FapC with a relative strong affinity but with possible effect from his-tag, and revealed potentially strong binders from a list of small molecules selected from a previous photofluorescence screening performed by Dr Kim Junyoung from the Chemical Bioimaging Laboratory, NUS. Optimization of the immobilization of FapC in the EnSpire® suggested conditions similar to those in the T200, while more optimization in the LigandTracer® is needed due to difficulty in immobilizing FapC. To conclude, signaling molecules and other small molecules were found to bind to FapC using Biacore T200™.