Figure_5.tif (3.72 MB)
Download file

CRM1 is retinoylated.

Download (0 kB)
posted on 24.06.2014, 03:33 by Sandeep Dave, Ravikanth Nanduri, Hedwin Kitdorlang Dkhar, Ella Bhagyaraj, Alka Rao, Pawan Gupta

(A) Cell extracts of adipocytes cells, transfected with FLAG-hCRM1, were subjected to in vitro retinoylation in presence of N-ethylmaleimide (NEM, 5 mM) or a nitrosating agent S-nitrosoglutathione (GSNO, 100 µM). The delipidated cell extract was later pulled down for FLAG and associated radioactivity was measured by liquid scintillation. (B) CRM1 mutants C528W and C585W were subjected to in vitro retinoylation and radioactivity was measured as mentioned above. Error bars show SD. * P<0.05 vs. controls. (C) Analysis of sequence contexts around cysteines in known retinoylated proteins (as listed in Table 1). A composite two-scale (at Y-axis) sequence weblogo presenting a general residue preference around cysteines (central) of known retinyolated proteins. The first half of the first bit of scale on Y-axis is expanded to clearly reflect the identity of residues around the central cysteine. The web logo here is only suggestive of general preferences and may not be statistically significant for the facts that so far no retinyolated sites are known to be validated experimentally and the number of proteins taken for analysis in the study is very small.