Dear All, I am working with a protein which is having an N-terminal unstructured region (approximate of length 40-60 amino acids) and C-terminal region which is having structured (possibly alpha helical region as probed by CD spectroscopy, Bioinformatic studies) property. The protein, in higher concentration (above 2mg/ml), forms various oligomeric structure (probed by Atomic force microscopy) resembling annular type aggregation (may be called as protofibril). when using various deletion mutants lacking different regions of N-terminus and C-terminus, we found that the C-terminal 45 residues is responsible and sufficient to make this annular type aggregation (like seen in alpha Synuclein, though much bigger is size than alpha Synuclein annular oligomers). We tried with our best to crystallize both the full length protein and the C-terminal 45 residues (which eventually has 37% sequence identity and 63% similarity [at Query coverage of 87%] with a putative UBA domain like structure of an archaeal protein). We tried various crystallization suites like PEG, PEG-II, Nucleix, pH clear etc to optimize the condition with no satisfaction at all. So, it would be a great help for me if I can get advises so as to troubleshot this problem. All advises are deeply appreciated.
Thank you, Best regards, Debasish Kumar Ghosh CSIR- Junior Research Fellow (PhD Scholar) C/o: Dr. Akash Ranjan Computational and Functional Genomics Group Centre for DNA Fingerprinting and Diagnostics Hyderabad, INDIA Email(s): dkgh...@cdfd.org.in, dgho...@gmail.com Telephone: 0091-9088334375 (M), 0091-40-24749396 (Lab) Lab URL: http://www.cdfd.org.in/labpages/computational_functional_genomics.html
