Department Bionanoscience
Principal investigator Dimphna Meijer
E-mail address d.h.m.meijer@tudelft.nl
Website www.dimphnameijerlab.org
Structural characterisation of FAM171A2 – FAM171B protein complex
Supervisor: Leanid Kresik, L.K.Kresik@tudelft.nl
In this project, we plan to perform structural biology studies of neuronal transmembrane proteins FAM171A2 and FAM171B from the FAM171 protein family. FAM171A2 and FAM171B are single-pass transmembrane proteins expressed in the brain interacting with each other. The existing literature provides limited information about the localization and functions of these proteins, and there is currently no available data about their structure. Using mammalian protein expression systems and protein purification techniques we will produce and purify single proteins and reconstitute FAM171-A2B complex in vitro. Next, we will utilise advanced electron microscopy technics such as negative stain electron microscopy and cryogenic electron microscopy for 3D molecular structure determination of the protein complex. Structural data will be complemented with other biophysical assays, such as Mass Photometry and Size-Exclusion Chromatography combined with Multi-Angle Light Scattering.
Techniques
- Cell culture
- Protein purification
- Size-Exclusion Chromatography
- Negative stain electron microscopy
- Cryogenic electron microscopy
- Mass Photometry
Unravelling the impact of patient CAMK2G mutations on protein activity and neuronal function
Supervisor: Angelica Casotto, a.c.casotto@tudelft.nl
Synaptic plasticity, a fundamental process in brain function, underpins the remarkable adaptability of the nervous system to external stimuli. In this process, the calcium/calmodulin-dependent protein kinase II (CAMK2) family emerges as a pivotal player. The CAMK2 family is a Ser/Thr protein kinase family consisting of four isozymes (CAMK2A, CAMK2B, CAMK2G, and CAMK2D). Recently, multiple de novo mutations in CAMK2G gene were identified among patients suffering from intellectual disability, highlighting the importance of CAMK2G in the brain. However, the mechanisms through which these mutations contribute to the onset of the disease remain poorly characterized. In this project we will use a combination of structural biology and biophysical assays to investigate the effects of these mutations on CAMK2G’s structure and activity.
Techniques
- Mammalian cell culture
- Protein purification
- Western blot
- Electron microscopy
- Molecular biology