Miguel,
I recommend the global phosphorylation method (JW Perich, 1991, Tetrahedron
Letters, 32:4033). I particularly like the recipe described in Methods in
Mol Biol, Vol 35: Peptide Synthesis Protocols, p. 195 (Ed. Pennington &
Dunn, 1994, Humana Press, Totowa, NJ).
Typically, you can expect reasonable results with either that recipe or
Novabiochem's prephosphorylated derivatives. Global is generally a lot
cheaper to perform, especially if you make a bunch, but it can be tricky.
Keep everything exquisitely dry for best results. I'd also generally
recommend the prephosphorylated derivatives for your first attempt, if you
only have one peptide to do. But this method is subject to rather severe
steric hindrance, both putting on the prephosphorylated Tyr, and acylating
it with the next AA or two.
Mark
-----Original Message-----
From: Microchemical Facility [mailto:mchem@socrates.Berkeley.EDU]
Sent: Tuesday, December 05, 2000 10:56 PM
To: Recipients of ABRF List
Subject: phosphotyrosine
Peptide synthesizers:
Could any of you 433 users suggest convenient/reliable method(s) for
preparing phosphorylated tyrosine containing peptides. Tech support
suggested using pre-dissolved PO3H2 fmoc tyrosine but this didn't give
desired peptide. Many thanks.
Miguel
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