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Please help: A proteolyzed product running bigger on an SDS gel?

Please help: A proteolyzed product running bigger on an SDS gel? - Protocols and Methods Forum

Please help: A proteolyzed product running bigger on an SDS gel? - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 05-29-2008, 05:19 AM
cv2135@columbia.edu
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Default Please help: A proteolyzed product running bigger on an SDS gel?



Hello
I'm trying to get to the bottom of a claim in my lab that a breakdown
product of our favorite protein actually runs at a HIGHER molecular
weight than the full-length product on a 10% SDS gel.
The breakdown product is supposedly 13 residues shorter at the C-terminus.


Our protein is:

-12.7 KD protein,

-very basic (with a PI of 9.42)

-binds RNA

-supposedly unstructured in the absence of its specific RNA ligand
(not generally present within E.coli)


Our protein runs at around 13-14 KD, and the supposed breakdown
product at 15KD

For those of you interested, the full-length protein sequence can be
found at :

[Only registered users see links. ]

(note first two residues, ML, represent a false start, and the NTD
methionine is cleaved in vivo, thus the length of the protein is
actually 108 AA)


Please , I'd appreciate it if anyone who can help me to understand an
explanation for this claim

Thanks so much
CV

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