Dear Friends,
I would like to chemically cleave a protein (133 aa) which has a (Asp) D - (Pro) P bond. This is method is working well,but I I would like to increase the digestion to 70% how to do it? also I would like to know what kind of modifications will take place because of this digestion

Can you eleborate on how you are chemically cleaving your peptide?

RB

R Bishop said:

Can you eleborate on how you are chemically cleaving your peptide? RB

1) I will add acetic acid to 13% (final concentration) and adjust the pH to 2.5 with pyridine and the concentration of the peptide will be 1mg/ ml. and proceed for cleavage by incubating for 3 days at 37Centigrade. In this manner I will be achieving about 20% cleavage.

2) If I increase the temperature to above 60C or 90C will there be any modifications (i.e deamidation or acetylation) to the protein? This I donot know? please kindly explain.

3) Does increase in temperature increases cleavage? Please kindly let me know how to increase the percentage cleavage.

Thank you
Sincerely yours
Dr.Murthy

You might try reading the protocols in this book

Basic Protein and Peptide Protocols - Humana Press

they have a chapter titled 'Enzymatic Methods for Cleaving Proteins'

I'll let you know if I figure anything else out

RB