Abstract

We studied the usefulness of genetic fusion to TELSAM polymers as an effective protein crystallization strategy. We observed novel properties in crystals of two TELSAM-target protein fusions. TELSAM as a crystallization chaperone shows rapid crystallization when it's fused to target proteins and possibly with a greater propensity. Some TELSAM-target fusions crystallized more rapidly than the same target protein alone. TELSAM-target proteins can be crystallized at relatively low protein concentrations such as 0.1 mg/mL. TELSAM requires no TELSAM polymers touching one another in the crystal lattice in order to form well-diffracting crystals. This lack of crystal contacts has not been observed in previously reported TELSAM crystal structures. Flexible TELSAM-target protein linkers can allow target proteins to find productive binding modes against the TELSAM polymer. This study tested TELSAM linker lengths varying by the number of glycines, such as 2xGly, 4xGly, 6xGly, 8xGly, and 10xGly. Only TELSAM fused to UBA with 2 and 4 glycine linkers were crystalized. TELSAM polymers can adjust their helical rise to allow fused target proteins to make productive crystal contacts, and fusion to TELSAM polymers increases avidity to stabilize weak inter-target protein crystal contacts. In conclusion, we report features of TELSAM-target protein crystal structures and outline future work needed to validate TELSAM as a crystallization chaperone and define the best practices for its use.

Degree

PhD

College and Department

Physical and Mathematical Sciences; Chemistry and Biochemistry

Rights

https://lib.byu.edu/about/copyright/

Date Submitted

2022-04-13

Document Type

Dissertation

Handle

http://hdl.lib.byu.edu/1877/etd12692

Keywords

TELSAM, Polymer forming Crystallization chaperone, Protein Crystallization, TNK1 UBA

Language

english

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