Affiliation:
1. Hefei National Laboratory for Physical Science at Microscale and School of Life Science, University of Science and Technology of China, Hefei, Anhui, China
Abstract
Background:
Kinetochores are the macromolecular protein complex that drives
chromosome segregation by interacting with centromeric DNA and spindle microtubules in
eukaryotes. Kinetochores in well studied eukaryotes bind DNA through widely conserved
components like Centromere Protein (CENP)-A and bind microtubules through the Ndc80
complex. However, unconventional type of kinetochore proteins (KKT1-20) were identified in
evolutionarily divergent kinetoplastid species such as Trypanosoma brucei (T. brucei), indicating
that chromosome segregation is driven by a distinct set of proteins. KKT proteins are comprised of
sequential α-helixes that tend to form coiled-coil structures, which will further lead to
polymerization and misfolding of proteins, resulting in the formation of inclusion bodies.
Results and Conclusion:
We expressed and purified the stable KKT proteins with Maltose Binding
Protein (MBP) fusion tag in E. coli or Protein A tag in Human Embryonic Kidney (HEK) 293T
cells. Furthermore, we identified interactions among KKT proteins using yeast two-hybrid system.
The study provides an important basis for further better understanding of the structure and function
of KKT proteins.
Funder
National Natural Science Foundation of China
Publisher
Bentham Science Publishers Ltd.
Subject
Biochemistry,General Medicine,Structural Biology
Cited by
2 articles.
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