[ccp4bb] MS method reference

Protein crystallography Programs for crystallography X-ray detectors
Basic tutorials:
Protein Peptide

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CCP4bb <-- 2007 <-- September 2007 <-- 07 September 2007

Previous message:
Subject: Coot and Density Fit Analysis
From: Andrew Gulick gulick {- at -} HWI {- dot -} BUFFALO {- dot -} EDU
Date: 2007-09-07

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Subject: Re: just how bad can phases be and still help
From: James Holton JMHolton {- at -} LBL {- dot -} GOV
Date: 2007-09-08

Subject: MS method reference
From: Jacob Keller j-keller2 {- at -} MD {- dot -} NORTHWESTERN {- dot -} EDU
Date: 2007-09-07

Hello All,

I was wondering whether anybody has a reference in which the following method is used:

Integral Membrane Protein (IMP) Topology by MS Fingerprinting. Express IMP, subject whole cells to
membrane-impermeable, membrane-soluble, and membrane-crossing, lysine-specific reagents to label
all free lysines in each compartment (i.e., extracellular, membrane-spanning, and intracellular
(and buried, which would not get labeled)). Then do rudimentary purification, run a gel, cut out
the band, run trypsinolysis-based fingerprinting, and compare the weights of the fragments with the
unlabelled. Those bands with a shift would have been exposed to the reagent in that experiment,
indicating that fragment/lysine's location with respect to the cell membrane.

I suspect that somebody must have thought of this previously...there is nothing new under the sun.

Jacob Keller

CCP4bb navigation

CCP4bb <-- 2007 <-- September 2007 <-- 07 September 2007

Previous message:
Subject: Coot and Density Fit Analysis
From: Andrew Gulick gulick {- at -} HWI {- dot -} BUFFALO {- dot -} EDU
Date: 2007-09-07

Next message:
Subject: Re: just how bad can phases be and still help
From: James Holton JMHolton {- at -} LBL {- dot -} GOV
Date: 2007-09-08