pLink 2 User Guide - ict.ac.cnpfind.ict.ac.cn/software/pLink/pLink2 User Guide.pdfpLink is an engine...
Transcript of pLink 2 User Guide - ict.ac.cnpfind.ict.ac.cn/software/pLink/pLink2 User Guide.pdfpLink is an engine...
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pFind Group
2020.01
pLink 2 User GuideVersion 2.3
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Contents
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❖Brief Introduction
❖Installation Requirement
❖Software Installation
❖Quick Start
❖Result Report
❖Appendix
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Brief Introduction
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❖pLink is an engine for cross-link peptide
identification, including peptides with chemical
crosslinking, endogenous crosslinking and
sumoylation.
❖The main form of peptide to be identified is inter-
linked peptide. Besides, it also supports other
forms, such as mono-linked peptides, loop-linked
peptides and so on.
❖The current version is 2.3 which is ~40 times
faster than pLink 1.
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Installation Requirement
❖Hardware
▪ CPU: 2.0 GHz or higher
▪ Memory: 4 GB or higher recommended
▪ Hard Disk: ~50 MB for software storage and an extra disk
space to store the results and temporary files
❖ Software
▪ Operating system: Windows 7/8/10, 64 bit version
▪ .NET Framework 4.5.2
▪ MSFileReader, 3.0 SP2 or below, both 32 bit and 64 bit
version, pLink 2 uses MSFileReader to access RAW files
▪ Java 8, 64 bit version, pLink 2 needs Java environment for
quantification
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Software Installation
❖ Double click pLink2.exe and install it in the chosen directory.
❖ Fill in personal information in the
pLink License Dialog.
❖ Copy and send the information to
[email protected] to get the license.
❖ Click “Import the license file” browse
and import the license.
❖ Finally, restart pLink 2.
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Main interface of pLink 2
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Create a new search task
❖ Click New…
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Create a new search task
❖ Fill the task name and browse the task location
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Import data
❖ You can choose MS data format and other data
preprocessing type in MS Data panel.
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Set search parameters
❖ A) Select flow type and cross linker
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Set search parameters
❖ B) Select and import database.
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Set search parameters
❖ B) Select and import database.
▪ Add contaminated proteins to the database if it doesn’t
contain them.
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Set search parameters
❖ B) Set the appropriate peptide mass range, peptide length
range, error range and modifications.
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Set search parameters
❖ C) Set the appropriate filter tolerance and FDR.
▪ As it is time-consuming to compute E-value, the default value
is unchecked.
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Set quantification parameters
❖ 15N labeling and Leiker labeling are supported if necessary
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Check parameters and run tasks
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Searching…
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Searching completed
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Contents of search results files
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Parameter file →
Web page result →
pQuant parameter file →
CSV results →
pLabel parameter files
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Web page result
❖ Click number with hyperlink to see the details.
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Web page result
❖ FDR Curve (Spectral Level).
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Web page result
❖ Precursor Error Distribution.
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CSV results
❖ Each peptide type has results in spectra, peptide and
site(protein) level.
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View search results
❖ Open *.plabel file with pLabel
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Appendix
❖ A) How to add a new linker?
1. Click Options→Meta Data Configuration
2. Select Linkers tab
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Appendix
❖ A) How to add a new linker?
3. Click Add, fill in linker information in the dialog
4. Click Update and then Save
5. Close pConfig window, the new linker will appear in
Linkers ListView
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Appendix
❖ A) Linker information needed
▪ Name: the name of the new linker.
▪ AlphaSites: the first cross-linked amino acid, parentheses “(” and
“)” denote the peptide N terminus and C terminus, respectively;
square brackets “[“ and “]” denote the protein N terminus and C
terminus, respectively.
▪ BetaSites: the second cross-linked amino acid, “(”, “)”, “[“, and “]”
denote the same as AlphaSites.
▪ LinkerMass: monoisotopic linker mass in inter/loop links.
▪ MonoMass: monoisotopic linker mass in mono links.
▪ LinkerComposition: linker composition in inter/loop links.
▪ MonoComposition: linker composition in mono links.
▪ LongMass: the longer mass in cleavable linker, 0 for uncleavable linker.
▪ ShortMass: the shorter mass in cleavable linker, 0 for uncleavable
linker.
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❖Thank you for using pLink 2!
❖If you have any questions, please contact
❖You can also post issues at GitHub for
discussion:
▪ https://github.com/pFindStudio/pLink2/issues
▪ how to post issues at GitHub?
• see http://pfind.ict.ac.cn/file/github.pdf
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