In article savidge at unb.ca, savidge at unb.ca ("Dr. R. A. Savidge") writes:
>A postdoc of mine isolated a protein from a pine tree by a series of
>fractionation steps including preparative IEF, SDS-PAGE and electroblotting
>to PVDF membrane, a band from the latter being N-terminal sequenced. A
>signal corresponding to about 5 pmoles was obtained. The sequence was
>NH2-DAHKSEVAHRFKDLGEEN and, according to the sequencer, this has 100%
>homology to human serum albumin as provided by the Swissprott data bank!
oops!
>I don't have direct access to Swissprott or other data banks. Can anyone
>confirm the homology interpretation?
yep, see gp:A03758 for an entry into the literature.
>We are certain that we did not use any type of serum albumin at any stage
>of protein purification. Sterile gloves were worn for the SDS and
>electroblotting procedures. The sequencer suggested that the gel may have
>been contaminated by blood, but that there would have had to have been a
>lot to obtain such a nice signal. We are certain that there were no cuts
>on fingers and that blood cannot be the explanation.
I don't suppose this piece of pine was used as an ice hockey stick before
it was sacrificed for science?
>I am going to repeat the purification and, funds permitting, will attempt
>another sequence. Does anyone have any other suggestions?
Imagine what the other guy is thinking: "How in the hell did I get a protein
from a pine tree in my sample of purified human serum albumin??!!!???"
>**********************************************************************
> Rod Savidge, PhD | E-mail: savidge at unb.ca> Faculty of Forestry and \|/
> Environmental Management \ | / Phone: (506) 453-4919
> University of New Brunswick _\/ | \/_
> Fredericton, NB CANADA \|/ Fax: (506) 453-3538
> E3B 6C2 |
---
TJ Murphy
Asst. Professor
Dept of Pharmacology
Emory University School of Medicine
