Development of a protein sensor for detecting histone acetylation
Abstract
H3K14ac (acetylation of lysine 14 of histone H3) is one of the most important epigenetic modifications found in cells. Aberrant changes in H3K14ac are commonly found in various types of cancers and neurological disorders. The current detection approaches for histone modifications, however, require either tedious sample pre-treatments or lack the quantitative accuracy required for biochemical and biomedical applications. In this study, we engineered a protein sensor using the amino acid sequences derived from human polybromo-1 (PB1). The protein sensor was conjugated to a fluorescent dye for sensitive detection of H3K14ac. Different detection conditions, such as additive concentrations and probe concentrations, were optimally selected by balancing signal strength (IRel) and signal-to-nosie ratio (SNR ). The protein sensor was verified using histone H3 peptides containing different H3K14 acetylation levels. The detection signal was found to be linearly dependent on H3K14acetylation levels ranging from 5 -100 %. The designed platform can be used for screening epgienetic drugs regulating H3K14 acetylation levels as well as monitoring H3K14 acetylation level of circulating nucleosomes for disease progression.
Degree
M.S.Ch.E.
Advisors
Yuan, Purdue University.
Subject Area
Chemical engineering|Oncology
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