RE: PepSyn: fluorescently labeled peptides

From: Singleton, David H (uunet!groton.pfizer.com!david_h_singleton@mail.uu.net)
Date: Tue Jan 11 2000 - 08:15:58 EST


IMHO:
1.) FMOC-Abz: No problem

FMOC-Tyr(mNO2)-OH: No problem with the free hydroxyl. You may notice salts
binding and releasing from it (conductivity will about double from previous
values) making your synthesis resin "pseudo ion-exchange" in character.
This is true with FMOC-Tyr(OH)-OH and the phenomenon disappears using UV
monitoring.

FMOC-Lys(DABCYL)-OH, or FMOC-Asp(EDANS): I wouldn't. If you've ever used
these in normal lab glassware; you'll know why. The dyes stain everything
and nothing ever decontaminates them again. You'll have pink or yellow
peptides forever.

2.) See above.

3.) We've cleaved doubly tagged peptides with reagent K many times and have
seen no problems. Follow the conventional wisdom re: Arg deprotection and
you'll be OK.

Hope this helps

David H. Singleton
Scientist
Pfizer Central Research
PO Box 8118-101
Eastern Point Road
Groton, CT 06340
-----Original Message-----
From: PaxtonR@immunex.com [mailto:PaxtonR@immunex.com]
Sent: Monday, January 10, 2000 8:19 PM
To: Recipients of ABRF List
Subject: PepSyn: fluorescently labeled peptides

We are interested in synthesizing a peptide on an ABI 433A for use in a
fluorescent quenching assay, and we have a few specific questions:

1. Are there any solubility problems with using cartridges loaded with
FMOC-Abz-OH, FMOC-Tyr(m-NO2)-OH, FMOC-Lys(DABCYL)-OH, or FMOC-Asp(EDANS)
-OH?

2. Are there any problems associated with the unprotected hydroxyl on the
side chain of FMOC-Tyr(m-NO2)-OH?

3. Does the use of any of these compounds in peptides require that the
cleavage conditions be modified?

Thanks for any help.
Ray Paxton



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