This density does not look at all like O-glycosylation. 
Best regards,
Herman


________________________________

        From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of 
Savvas Savvides
        Sent: Wednesday, November 24, 2010 2:48 PM
        To: CCP4BB@JISCMAIL.AC.UK
        Subject: Re: [ccp4bb] Strange density on Serine oxygen.
        
        
        Hi Vinson 
        is O-glycosylation possible at all given the origin of the protein and 
the expression system used? 
        best 
        Savvas
        
        
        



        On 24 Nov 2010, at 14:42, Vinson LIANG wrote:


                
                Dear Savas and all,
                 
                Thank you very much for all your quick suggestions.
                 
                I have tried Tyr and it turns out to fit the density very well. 
I will have the protein sequenced again to see if it is wrong sequece or 
O-linked glycosylation.
                 
                I'll let you know if it turns out to be O-linked glycosylation.
                 
                All the best,
                 
                Vinson 
                
                
________________________________

                发件人: Savvas Savvides <savvas.savvi...@ugent.be>
                收件人: Vinson LIANG <lwg_conrad_1...@yahoo.com.cn>
                抄 送: CCP4BB@JISCMAIL.AC.UK
                发送日期: 2010/11/24 (周三) 9:27:31 下午
                主 题: Re: [ccp4bb] Strange density on Serine oxygen.
                
                Hi Vinson 
                Beyond the possibility for another type of residue as already 
suggested by Phil and Mark, there is also the possibility of O-linked 
glycosylation of the serine and threonine, if your protein undergoes such 
post-translational modification and it has been expressed via an expression 
system that processes the protein in that way.
                Ser/Thr tandems are well known targets for O-glycosylation 
(http://www.cbs.dtu.dk/databases/OGLYCBASE/).

                best regards
                Savvas

                
                ----
                Savvas Savvides
                Unit for Structural Biology @ L-ProBE
                Ghent University
                K.L. Ledeganckstraat 35, 9000 Ghent, Belgium
                Ph. +32  (0)472 928 519 http://www.LProBE.ugent.be/xray.html



                On 24 Nov 2010, at 13:10, Vinson LIANG wrote:


                        
                        Dear all,
                         
                        I'm refining a structure and find some strange triangle 
density on the oxygen of Ser and Thr at the C terminus. One picture of the 
strange density is attached here. Could anyone please give me some suggestions 
on what this could be?
                         
                        The buffer used during purification is PBS, Tris and 
NaCl. And crystallization condition contains PEG3,350 and Mg(NO3)2.
                         
                        Thank you all in advance for any suggestion.
                         
                        Best,
                         
                        Vinson Liang
                         
                         

                         <triangle_density.gif>



                 

                 


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