Kinase activated by Glycerol

Vinnie Ossipow at sc2a.unige.ch
Sat May 13 12:29:45 EST 1995


>Alexandre Maucuer
>INSERM U153                                     phone (33) 1 43 37 17 23
>17, rue du Fer a Moulin, 75005 Paris, France    email maucuer at citi2.fr
From: Ossipow at sc2a.unige.ch (Vinnie)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: Kinase activated by Glycerol
References: <3odko6$een at bisance.citi2.fr>
Organization: Universite de Geneve
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In article <3odko6$een at bisance.citi2.fr>, maucuer at bisance.citi2.fr (Maucuer) says:
>
>hello
>
>I produced a GST-Kinase fusion protein. When i tested its
>autophosphorylation after elution with glutathione and 
>dialysis against kinase buffer, i observed a dramatic
>enhancememt by adding 25% glycerol in the reaction.
>
>Does anyone has experience about such effect ?
>
>The GST-Kinase was contaminated by seemingly DnaK which
>also seems to autophosphorylate.
>
>Does anyone know about substrate(s) and specificity of
>phosphorylation by DnaK ?
>
>Thanks.
>
>Alex Maucuer
>-- 
>Alexandre Maucuer
>INSERM U153                                     phone (33) 1 43 37 17 23
>17, rue du Fer a Moulin, 75005 Paris, France    email maucuer at citi2.fr
I have also noticed autophosphorylation activity by a 
70 KDa protein from E. coli that co-purifies with
GST. I don't know if it is DnaK, neither its substrate 
specificity but I know for sure its MW because I have
detected it by an "in gel phosphorylation assay". What
 about you ?

		see you 



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