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44 changes: 30 additions & 14 deletions vignettes/MSstatsPTM_LabelFree_Workflow.Rmd
Original file line number Diff line number Diff line change
Expand Up @@ -57,19 +57,6 @@ for your tool in `MSstatsPTM`, you can alternatively leverage converters from
base `MSstats`. If using converters from `MSstats` note they will need to be
run both on the global protein and PTM datasets.

You might notice a FASTA file is also needed for some converters. This
FASTA file can be obtained by querying
[Uniprot](https://www.uniprot.org/id-mapping) with all of the protein
IDs present in your PTM dataset. The FASTA file is a necessary input
because some tools (e.g. MaxQuant) do not report the specific amino acid
that is modified relative to the whole *protein* sequence. Rather, they
report the specific amino acid relative to the reported *peptide*. This
distinction is important because modifications like phosphorylation,
methylation, or acetylation often have specific roles depending on where
they occur within the full-length protein. With the help of a FASTA
file, MSstatsPTM can determine the specific amino acid that is modified
in the context of the whole protein sequence.

Please note for the PTM dataset, both the protein and modification site (or
peptide), must be added into the `ProteinName` column. This allows for the
package to summarize to the peptide level, and avoid the off chance there are
Expand Down Expand Up @@ -240,7 +227,36 @@ Experiments can be acquired with label-free labeling methods.
`MSstatsPTM` includes a dedicated converter for Progenesis output.
Experiments can be acquired with label-free labeling methods.

#### 1.1.7 Additional tools
#### 1.1.7 FASTA File

You might notice a FASTA file is also needed for some converters. This
FASTA file can be obtained by querying
[Uniprot](https://www.uniprot.org/id-mapping) with all of the protein
IDs present in your PTM dataset.

Follow these steps to download a FASTA file from UniProt:

1. **Prepare your protein IDs** Make a list of your protein IDs (e.g., UniProt accessions like P31749, Q9Y243, etc.). You can copy-paste them into the UniProt tool.
2. **Go to the UniProt ID mapping tool** Open the UniProt ID mapping page: https://www.uniprot.org/id-mapping
3. **Select input database** In the "From" dropdown, choose: UniProtKB AC/ID
4. **Select output database** In the "To" dropdown:
+ If isoform-specific data is needed, select: UniProtKB/Swiss-Prot
+ Otherwise, select: UniProtKB
5. **Submit your IDs** Paste your protein IDs into the text box and click "Submit".
6. **Download the FASTA file** Once the mapping is complete, you’ll see a results page.
+ Click the "Download" button on the top of the table.
+ Select "FASTA (canonical)" and click download

The FASTA file is a necessary input because some tools (e.g. MaxQuant) do not
report the specific amino acid that is modified relative to the whole *protein*
sequence. Rather, they report the specific amino acid relative to the reported
*peptide*. This distinction is important because modifications like phosphorylation,
methylation, or acetylation often have specific roles depending on where
they occur within the full-length protein. With the help of a FASTA
file, MSstatsPTM can determine the specific amino acid that is modified
in the context of the whole protein sequence.

#### 1.1.8 Additional tools

If there is not a dedicated `MSstatsPTM` converter for a processing tool,
the existing converters in `MSstats` and `MSstatsTMT` converters can be used as
Expand Down
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