I think it completely depends on the protein: in some proteins, they are required for folding; in some (eg Fcs), they are not required for folding but for function); in some, some of them are required and others not; in some they are required _during_ folding, but afterwards they can be removed without affecting apparent stability. We had that experience with some UDP glycosyl transferases.
Adrian > On 12 Apr 2017, at 07:52, Jan Dohnalek <dohnalek...@gmail.com> wrote: > > We had experience with a relatively small glycoprotein - when glycosylation > sites were deleted, solubility went drastically down - we could not express > soluble any more. Back to eukaryotic expression system which worked. > > So may be you were really lucky. > > Jan > > > On Tue, Apr 11, 2017 at 10:34 PM, Bernhard Rupp <hofkristall...@gmail.com > <mailto:hofkristall...@gmail.com>> wrote: > Hi Fellows, > > > > a humble question for our glyco-expressionists: > > > > I have mutated out the Asns of the N-glycoslation consensus sites for Asp > > (Asp simply because the PNGaseF treated protein stays stable so I thought > that might be a good guess) > > and indeed the unglycosilated mutant expresses well and gets secreted as > planned. > > > > But rumor has it that glycoproteins that are mutated to non-glyc often are > not processed correctly and > > that we had just dumb luck. > > > > May I poll the educated opinion of the erudite here? > > > > Cheers, BR > > > > ------------------------------------------------------ > > Bernhard Rupp > > Crystallographiae Vindicis Militum Ordo > > http://www.hofkristallamt.org/ <http://www.hofkristallamt.org/> > b...@hofkristallamt.org <mailto:b...@hofkristallamt.org> > +1 925 209 7429 <tel:(925)%20209-7429> > +43 767 571 0536 <tel:+43%207675%20710536> > ------------------------------------------------------ > > :(){ :|: & };: > > ------------------------------------------------------ > > > > > > > -- > Jan Dohnalek, Ph.D > Institute of Biotechnology > Academy of Sciences of the Czech Republic > Biocev > Prumyslova 595 > 252 50 Vestec near Prague > Czech Republic > > Tel. +420 325 873 758
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