1. Metal-binding proteins:
Metallothioneins (MT) are a class of metal-binding proteins that are involved
in heavy metal detoxification and regulation in a wide range of organisms
from fungi to mammalian systems. We are interested in engineering metal scavengers
that can be used either in situ or in free environment for phytoremediation
of heavy-metal polluted soils and for metal recycling using metallothionein
metal centers as models. The results can also be used to engineer heavy-metal
resistance in other economically important plants such as tomato, corn, and
soybean.
We identified a novel metallothionein gene in pasta wheat (T.durum) and are
in the process of cloning the mt for overexpression of the recombinant protein
in E.coli. T.durum MT will be purified for structural studies using X-ray
solution scattering and X-ray crystallography. A recombinant GFP-MT fusion
protein will also be expressed for easy localization of the protein in transgenic
plants.
People working on this project: K. Bilecen and Ü. Öztürk
Collaboration: Dr. İ. Çakmak, Sabancı University and Dr. M. Koch EMBL Hamburg
Outstation.
Grant: Supported by Julich-TUBITAK bilateral program.
2. G-proteins:
Heterotrimeric G-proteins are involved in signal transduction in different
species. We are interested in understanding the role of G-proteins in pathogen
response in plants. Currently we are in the process of cloning the alpha
and gamma subunits of A. thaliana G-protein. The recombinant proteins will
be overexpressed in E. coli. Structure determination will be carried out
using X-ray crystallography and protein-protein interactions will be studied
using X-ray solution scattering on reconstructed complexes. We also aim to
construct recombinant subunits with chromophores for detecting the G-protein
complexes in situ.
People: S. Bakkal, Ç. Seçkin and M. Şahin
Collaboration: Dr. B. Kan Marmara University and K. Brown, Imperial College, UK.
3. Development of novel materials and systems for biological assays:
Here we aim to engineer a novel flexible gene expression system that can
be used for immobilisation and detecting functional activity of proteins,
enzymes etc. Such systems could serve as a basis for single step assay-detection
kits. The expression system under construction is derived from pETM-11 of
EMBL and will use built-in additional components e.g. chromophores for detection
and adaptors for immobilisation.
People: E. Şahin
Collaboration: Dr. A. Taralp, Sabanci University.
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