Immunological Strategies to Study GRP170 in Caenorhabditis elegans

Department Chair

Martha Skerrett, Ph.D.

Author

Raven P. Baxter-Christian, State University of New York College at Buffalo - Buffalo State CollegeFollow

Date of Award

8-2017

Access Control

Open Access

Degree Name

Biology, M.A.

Department

Biology Department

Advisor

Gregory J. Wadsworth, Ph.D.

Department Home page

http://biology.buffalostate.edu/

First Reader

Gregory J. Wadsworth, Ph.D.

Second Reader

Derek L. Beahm, Ph.D.

Third Reader

Daniel L. Potts, Ph.D.

Abstract

Characterization of the ER protein folding chaperone GRP170 of Caenorhabditis elegans could be greatly facilitated by an antibody which recognized the chaperone. Antibodies have not been raised against nematode GRP170. Groups have prepared polyclonal antibodies against vertebrate forms of GRP170. My thesis goal was to investigate whether anti-vertebrate GRP170 antibodies can recognize the nematode homologue on a standard Western Blot assay. Sequences of antigens that groups used to generate anti-vertebrate GRP170 antibodies were analyzed. Peptides used by Ruan et al. (2013), which correspond to regions of human GRP170, shared greatest sequence similarity with nematode GRP170. I investigated whether the Ruan GRP170 antibody recognized nematode protein on Western Blots. I extracted mouse liver proteins and whole worm proteins from C. elegans, separated proteins by SDS-PAGE, transferred proteins to a PVDF membrane, and probed membranes with the Ruan anti-human GRP170 antibody. Although this antibody did recognize a high molecular weight mouse protein, it did not bind to an equivalent high molecular weight C. elegans protein. This antibody would have limited utility for studies of nematode GRP170. Isoform-specific antibodies generated against C. elegans GRP170 proteins would be valuable tools to differentiate the functions of the two GRP170 isoforms. I analyzed amino acid sequences of the two GRP170 isoforms of C. elegans. I identified two peptides in a highly-diverged region of C. elegans GRP170, possible isoform specific antigens. Antibodies raised against these peptides could show isoform specific recognition and be useful for characterizing the two-gene, two-protein GRP170 system in C. elegans.

Recommended Citation

Baxter-Christian, Raven P., "Immunological Strategies to Study GRP170 in Caenorhabditis elegans" (2017). Biology Theses. 29.
https://digitalcommons.buffalostate.edu/biology_theses/29