Hi  all

I am trying to use the latest version TPP (4.3.1) to run some cICAT
data I collected a while ago. All these cICAT data have been searched
by Mascot. I also have run the tutorial data (raft with older version
of ICAT labeling) on my computer. Since the tutorial data were
searched by Sequest, so I looked up spctools-discuss for some
discussion about Mascot data, but I still have some Mascot related
questions. I am currently using Mascot server 2.2.04.

1.      If I have a Mascot.dat file, then can I directly start with
“Analysis Pipeline (Mascot)/ pepXML” (don’t need any database search
under TPP) ?

In “pepXML/ 2. Specifiy Database Used in Mascot Search” means that I
need to put the EXACT database I used for this searche? For instance,
my searches were done by against database SwissProt 55.6, so do I need
to choose the SwissProt_55.6.fasta file here? The current SwissPort
version is 57.12 in our lab, can I use SwissProt_57.12.fasta file for
this step? If not, where can I download the older version SwissProt
database? I tried, sounds silly, but I didn’t find any archive for
downloading the older SwissProt database .fasta files.

Anyway, I searched my data again under current SwissProt database
(57.12), but still didn’t get it worked. Here is the error massage I
got (more details see attachment).

c:/Inetpub/wwwroot/ISB/data/SSM4-1/F030337.pep.xml [ Unreadable! ]

command "c:\Inetpub\tpp-bin\Mascot2XML c:/Inetpub/wwwroot/ISB/data/
SSM4-1/F030337.dat -Dc:/Inetpub/wwwroot/ISB/data/SwissProt_57.12.fasta
-Etrypsin" failed: Operation not permitted
Command FAILED


2.      I also tried to use Mascot.xml and .pepxml files to directly start
with “Analyze Peptide”. The .xml gave no result (can’t open the pepXML
Viewer) and I couldn’t convert it into pepXML format under pepXML in
TPP.

If I used .pepxml file from Mascot, it gave the results but not
including any XPRESS and ASAPRatio result. From the online discussion,
it seems the Mascot.dat files work better than xml/pepxml files in
TPP. If I can get 1st problem resolved, then I don’t need to worry
about this 2nd problem.

Detailed error messages please see attachment.

Many thanks for any assistance!

Fay

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