Re: [ccp4bb]: off-topic: deglycosylation

[Anastassis Perrakis]

You can express it in GnTI(i) HEK293S cells. See  Structure and function in rhodopsin: high-level _expression_ of rhodopsin with restricted and homogeneous N-glycosylation by a tetracycline-inducible N-acetylglucosaminyltransferase I-negative HEK293S stable mammalian cell line. GnTI(-) HEK293S http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12370423&query_hl=7&itool=pubmed_docsum A. On Feb 2, 2006, at 13:58, Paul McEwan wrote: ***  For details on how to be removed from this list visit the  *** ***          CCP4 home page http://www.ccp4.ac.uk         *** I'm currently working with a glycoprotein and am having great difficulty deglycosylating it for crystallisation. I can completely remove the sugars very easily when the protein is denatured (not much good when you're wanting to crystalise it), but seems completely resistant to treatment with endo H or pngase F when done under native conditions. Any hints on getting this to work would be greatly appreciated.  ################################## Dr Paul A. McEwan Protein Crystallography Group Centre for Biomolecular Science Clifton Boulevard University of Nottingham Nottingham NG7 2RD Tel: 0115 8468009 http://www.nottingham.ac.uk/~pazxtal ################################## This message has been checked for viruses but the contents of an attachment may still contain software viruses, which could damage your computer system: you are advised to perform your own checks. Email communications with the University of Nottingham may be monitored as permitted by UK legislation.