Biotinylation of N-terminus only
robison1 at husc10.harvard.edu
Thu Feb 18 00:06:31 EST 1993
tshin at husc8.harvard.edu (Tae Shin) writes:
>I am interested in labelling a protein (not an antibody) using N-hydroxy-
>succinimide biotin. The standard protocol requires this labelling be done at
>pH9, which should label all of the accesible lysine sidechains and the
>N-terminus. I have heard that by lowering the pH (to about 6) one should be
>able to label the N-terminus selectively. Has anyone had any experience with
>this? Any comments would be welcome. Thanks.
>tshin at husc8.harvard.edu
I don't know if anyone has done this with biotin, but you should
check out the following reference by our own Dan Jay:
Jay, DG. 1984. JBC 259:15572-15578.
A general procedure for the end labeling of proteins and positioning
of amino acids in the sequence.
In brief, the scheme is to perform a single round of Edman degredation
on the protein. This results in a protein which is missing the N-terminal
amino acid, but with all its lysines blocked with the Edman group. You
can then proceed to label the remaining N-terminus specifically.
Department of Cellular and Developmental Biology
Department of Genetics / HHMI
robison at ribo.harvard.edu
More information about the Methods