UROP Project

Investigating phosphorylation-dependent α-actinin-2 and CaMKII interactions in cultured human cells

Bioengineering and biomedical engineering, Biological Sciences, Biochemistry, Cell Biology, Cardiovascular Biology, Signal Transduction, Protein-Protein Interactions, Cytoskeletal Proteins, Fluorescence Microscopy, Heart Failure

Research Mentor: Dr. Helene Tigro, She/Her
Department, College, Affiliation: Florida State University, Education, Health, and Human Sciences
Contact Email: ht24@fsu.edu
Research Assistant Supervisor (if different from mentor): Dr. Christopher Solis He/His
Research Assistant Supervisor Email: csolis@fsu.edu
Faculty Collaborators:
Faculty Collaborators Email:

Looking for Research Assistants: Yes
Number of Research Assistants: 1
Relevant Majors: Biochemistry, Neuroscience, Chemistry, Biology, Computational Biology, Engineering
Project Location: On FSU Main Campus
Research Assistant Transportation Required:
Remote or In-person: In-person
Approximate Weekly Hours: 10 hours, During business hours
Roundtable Times and Zoom Link:

• Day: Wednesday, September 2
  Start Time: 1:00
  End Time: 2:30
  Zoom Link: https://fsu.zoom.us/j/98918911622

Project Description

α-Actinin-2 (ACTN2) is a major structural protein of the cardiac sarcomere that crosslinks filamentous actin (F-actin) and anchors signaling proteins at the Z-disc. In addition to its structural role, α-actinin-2 functions as a signaling scaffold that coordinates mechanical and biochemical responses to cellular stress. Recent studies have identified direct interactions between α-actinin-2 and calcium/calmodulin-dependent protein kinase II (CaMKII), a key regulator of cardiac remodeling, calcium handling, and stress signaling. Dysregulation of CaMKII signaling contributes to the development of heart failure and cardiomyopathy.
Our laboratory studies how phosphorylation of the α-actinin-2 actin-binding domain regulates protein structure, actin binding, and signaling interactions. Preliminary computational and biochemical studies suggest that phosphorylation-induced conformational remodeling of the α-actinin-2 actin-binding domain may expose signaling interfaces that influence interactions with CaMKII. However, the mechanisms by which phosphorylation regulates α-actinin-2-CaMKII interactions remain poorly understood.
This project will investigate the cellular localization and interaction of α-actinin-2 and CaMKII using cultured human cells. Students will gain hands-on experience in mammalian cell culture, transient transfection, fluorescence microscopy, and quantitative image analysis while contributing to ongoing studies examining the relationship between cytoskeletal organization and cellular signaling. Understanding how α-actinin-2 and CaMKII interact may provide insight into mechanisms that regulate cardiac adaptation and remodeling in health and disease.

Research Tasks: The student will conduct a literature review on α-actinin-2, CaMKII signaling, and regulation of the cardiac cytoskeleton. Research activities will include routine maintenance and passaging of HEK293T cells, preparation and amplification of plasmid DNA, and transient transfection of fluorescently tagged α-actinin-2 and CaMKII constructs. The student will monitor transfection efficiency and protein expression using fluorescence microscopy and assist in optimizing experimental conditions. Imaging studies will be performed to evaluate the subcellular localization and colocalization of α-actinin-2 and CaMKII. Quantitative image analysis will be carried out using ImageJ/FIJI to measure fluorescence intensity, protein distribution, and colocalization parameters. The student may also assist with immunoblotting, protein expression analysis, data interpretation, statistical analysis, and graphical presentation of results.

Skills that research assistant(s) may need: Good theoretical understanding of biology and chemistry. Interest in cell biology, protein biochemistry, cardiovascular biology, and molecular signaling pathways. Experience with laboratory techniques is helpful but not required. Experience with fluorescence microscopy, image analysis software, molecular biology methods, or programming (Python, R, MATLAB) is beneficial but not required. Strong organizational, time-management, and communication skills, along with the ability to work independently and as part of a team, are highly valued.