Re: PTH-Cys-CAM

Rod Levine (rlevine@nih.gov)
Wed, 09 Jun 1999 08:49:41 -0400

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At 02:57 PM 6/9/1999 +0800, leefeng wrote:

> I have modified some of cysteine residues of a polypeptide using IAA.
>I want to know the position of labled cysteine residues by sequence
>analysis. But there is no PTH-Cys-CAM in my PTH-AA standard. Could you
>tell me the position of PTH-Cys-CAM on PTH-AA standard chromatogram?

On our Hewlett-Packard sequencer, it runs between Gly and His -- nicely
separated from both.

If you use iodoacetic acid to generate the CM-Cys-PTH, it runs just after Glu
-- and will not fully separate from it, but the later time of the peak is quite
reproducible. We prefer using iodoacetamide to avoid ambiguity.

Rod Levine

NIH
Bldg 3, Room 106 MSC 0320
Bethesda, MD 20892-0320

email: rlevine@nih.gov
voice: 1 (301) 496-2310
fax: 1 (301) 496-0599
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At 02:57 PM 6/9/1999 +0800, leefeng wrote:

>  I have modified some of cysteine residues of a polypeptide using IAA.
>I want to know the position of labled cysteine residues by sequence
>analysis. But there is no PTH-Cys-CAM in my PTH-AA standard. Could you
>tell me the position of PTH-Cys-CAM on PTH-AA standard chromatogram?


On our Hewlett-Packard sequencer, it runs between Gly and His -- nicely separated from both.

If you use iodoacetic acid to generate the CM-Cys-PTH, it runs just after Glu -- and will not fully separate from it, but the later time of the peak is quite reproducible.  We prefer using iodoacetamide to avoid ambiguity.

Rod Levine


NIH
Bldg 3, Room 106 MSC 0320
Bethesda, MD 20892-0320

email: rlevine@nih.gov
voice: 1 (301) 496-2310
fax:   1 (301) 496-0599

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