The C-terminal residue can be a bit tricky sometimes. In general, anything
that is hydrophilic will be retained by biobrene and you should see it OK.
In fact, residues such as Arg can end up repeating themselves through
inefficient extraction. However, very hydrophobic residues will be washed
out to varying degrees and you may see them as a residue with greatly
reduced yield. With synthetic peptides this is not generally an issue as
you are able to load high pmol amounts initially if necessary. We did have
a peptide though with a C-terminal sequence of FIL and we never did manage
to see the last two residues with any confidence, so we used MS-MS instead.
If you have at least linear maldi mass analysis capability, then the
C-terminal residue can be deduced by comparing mass to sequence (Q/K, I/L
excepted), or by using C-terminal sequencing kit (PerSeptive).
Len
>>Hi everybody,
>>
>>Can anyone tell me if they have had any problems sequencing the c-terminal
>>AA of a synthesized peptide? Does the final AA still stick to the Biobrene
>>filter or can it get washed off resulting in very low or no yields?
>>
>>Thanks in advance
>>
>>Amanda
>>
*********************************************************************
Dr Len C. Packman
Assistant Director of Research
Protein and Nucleic Acid Chemistry Facility
Department of Biochemistry
University of Cambridge
80 Tennis Court Road
Old Addenbrookes Site
Cambridge, CB2 1GA, UK
Tel: +44 (1223) 333639
FAX: +44 (1223) 766002
e-mail: lcp2@mole.bio.cam.ac.uk
Visit my WWW page at http://www.bio.cam.ac.uk/proj/adr/PNAC/pnac.html