Example of Process Workflow for Pyruvate Kinase

(MOWSE results at bottom of the page):

Data generated by Leila Choe.

Spots can be excised from fluorescently stained gels using a KBiosystems KCore. The resulting spots are digested robotically with trypsin using a Genomic Solutions ProGest. The digests are cleaned by Ziptip using a Packard Multiprobe IIEX. Peptides and matrix are spotted automatically to 4700 target plates using the Applied Biosystems Symbiot XVI. This process is depicted schematically here.

A Peptide Mass Fingerprint from a Pseudomonas syringae protein.

Peaks used for MSMS identified with an arrow. We can routinely collect multiple MSMS runs from a single sample.

MSMS of m/z 2762 from PMF:

MSMS of m/z 1258 from PMF:

MSMS of m/z 1827 from PMF:

MSMS of m/z 2925 from PMF:

MSMS of m/z 1599 from PMF:

When this data is submitted for a search, our results look like this...

Note that some of the links are clickable and allow you to view some of the MSMS data and click through to NCBInr searches for hits against pyruvate kinase. Resizing MSMS plot under queries will fail.

When we process the data using Genevieve DeClerck's scripts to align the observed peptides onto the syringae genome we get this type of information... From here you can also get to NCBInr searches of this sample.

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