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Identifying Protein Partners of CFAP69

Undergraduate #42
Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology

Lewis Bennett - University of the Virgin Islands
Co-Author(s): Frederick Dong, The Johns Hopkins University, Baltimore, MD; Haiqing Zhao, Ph.D, The Johns Hopkins University, Baltimore, MD



Reproduction is fundamental to the continuation of life. Many of the processes in reproduction including spermatogenesis, the process of sperm cell development, have yet to be fully understood. Cilia flagella associated protein 69 (Cfap69) is an evolutionarily conserved, yet poorly characterized protein. Our laboratory recently found that CFAP69 mutant male mice had defective sperm, suggesting CFAP69 is required for spermatogenesis. Analysis of the protein sequence indicates that a large portion of the CFAP69 contains an ARMADILLO domain, which typically mediates protein-to-protein interactions. In order to understand the role and mechanism of CFAP69 in spermatogenesis, we attempt to identify its interaction partners. CFAP69 and the c-terminal portion of the CFAP69 were expressed in E. coli as a GST fusion proteins, and the purified fusion proteins were used as baits to capture any interacting proteins from testis lysates. CFAP69 and its interaction partners would be eluted from gluthathione columns, and its interaction partners identified by mass spectrometry. Further analysis of these proteins will help to clarify CFAP69’s involvement in reproduction.

Not Submitted

Funder Acknowledgement(s): MARC Grant 5T34GM008422 National Science Foundation REU site grant

Faculty Advisor: Haiqing Zhao, Ph.D, hzhao@jhu.edu

Role: Expressed the c terminal portion of CFAP69 as a GST fusion protein in E. coli. Purified the fusion proteins and performed a gst pull down assay to capture any interaction proteins of CFAP69 from mouse testis lysates.

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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