RE: LysC autolytic peptides

Jelle Lahnstein (jlahnste@waite.adelaide.edu.au)
Mon, 17 Aug 1998 12:36:41 +0930 (CST)

In addition to the Boehringer and Wako LysC endoproteinases, the Promega
version is also commonly used (around here anyway). Their source is
Pseudomonas aeruginosa. I've always had good results with it under a wide
range of conditions, and it's a lot cheaper than from Boehringer.

Jelle

____________________________________________________
Jelle Lahnstein
Nucleic acid and Protein Chemistry Unit
Department of Plant Science
Waite Campus, University of Adelaide
Glen Osmond, 5064, Australia
Tel : 61 - 8 - 83037260
Fax : 61 - 8 - 83037130
Internet : http://www.waite.adelaide.edu.au/NAPCU/


On Fri, 14 Aug 1998, Edmunds, Tim wrote:

> A word of caution.
>
> There are two proteases commonly used for cleavage at Lys residues.
> Wako sells Lysyl Endopeptidase from Achromobacter lyticus E.C.
> 3.4.21.50. The enzyme from Boehringer is Endoproteinase Lys-C from
> Lysobacter enzymogenes E.C.3.4.99.30. Unfortunately, for convenience,
> both of these proteinases are often called Lys-C. When looking for
> autolytic fragments it is obviously important to make sure you are
> trying to match masses to the correct sequence.
>
> I know the general consensus has been that the Wako enzyme gives more
> reproducible results but we have not found that to be true for the
> proteins we have digested. We switched from the Wako enzyme to the
> Boehringer sequencing grade enzyme many years ago due to a high
> background of autolytic fragments and incomplete digestion products
> obtained with the Wako enzyme. Obviously there is a significant cost
> difference between the Boehringer sequencing grade enzyme and the Wako
> enzyme and our observations may not hold up for the Boehringer bulk
> Lys-C.
>
> Tim
>
> Tim Edmunds
> Director
> Structural Protein Chemistry
> Genzyme Corporation
> P.O Box 9322
> Framingham
> MA 01701-9322
>
> Tel (508) 270 2260
> Fax (508)872 9080
> EMail tedmunds@genzyme.com
>
>