Re: PepSyn: Difficult Sequence

Michael Pereckas (msp@ace.biochem.mcw.edu)
Mon, 9 Feb 1998 17:01:35 -0600

On Mon, Feb 09, 1998 at 12:09:54PM -0600, procor@smtpgate.bcsew.edu wrote:

We switched from the OPFp esters to HATU activation after some
much-improved results on a peptide that we were having trouble with.
That might help. This sequence actually doesn't look particularly
hydrophobic. I've synthesized much more hydrophobic peptides without
any special problems.

> I am having a problem synthesizing a peptide with the sequence
> PITKYIMTCMSADLEVVT. I am using std FMOC chemsitry with OPfp esters,
> 20% Pip as the deblock, DMA as the solvent and PEG-PS as the starting
> resin. I am using a 9050 instrument. I dried the resin and cleaved in
> 92.5 % TFA, 2.5% TIS, 2.5% ethanethiol for 2 hrs. I precipitated the
> peptide in fresh ether and washed it 3x. The pellet was dried on a
> lyophilizer. The resulting pellet was very granular. I tested the
> crude peptide using RPHPLC, 5-60% ACN and saw no clear peak. I sent
> the crude out for ES MS analysis and the tracing showed alot of noise
> but no target peptide. I know this sequence is very hydrophobic and I
> may have to change my purification strategy but I don't feel like I
> even have the peptide on the resin yet. Any sugesstions would be
> appreciated.

-- 
Michael Pereckas  //  msp@mcw.edu  //  Michael.Pereckas@mixcom.com
Protein and Nucleic Acid Shared Facility, Medical College of Wisconsin
Research Technologist and sysadmin/webmaster of www.biochem.mcw.edu