Vermont Genetics Network Outreach Proteomics Module - PowerPoint PPT Presentation

Loading...

PPT – Vermont Genetics Network Outreach Proteomics Module PowerPoint presentation | free to download - id: 70f55f-MWUwZ



Loading


The Adobe Flash plugin is needed to view this content

Get the plugin now

View by Category
About This Presentation
Title:

Vermont Genetics Network Outreach Proteomics Module

Description:

Vermont Genetics Network Outreach Proteomics Module Protein Mass Spectrometry: Theory and Application Prepared by Bryan A. Ballif, Ph.D. Two Essential Partner Tools ... – PowerPoint PPT presentation

Number of Views:77
Avg rating:3.0/5.0
Slides: 15
Provided by: Coop51
Learn more at: http://www.uvm.edu
Category:

less

Write a Comment
User Comments (0)
Transcript and Presenter's Notes

Title: Vermont Genetics Network Outreach Proteomics Module


1
Vermont Genetics Network OutreachProteomics
Module
Protein Mass Spectrometry Theory and
Application Prepared by Bryan A. Ballif, Ph.D.
2
Two Essential Partner Tools in Proteomics
3
First Things First -- Know Your Goal !
? There are nearly as many mass spectrometry
methods as there are mass spectrometry
projects.
? Your goal determines your methods which
determine your outcomes.
4
The Four Most Common Protein Mass Spectrometry
Projects (Observe How Each Project Has A Distinct
Goal / Method !!)
5
Typical Protein Mass Spectrometry Work Flow
Protein Preparation / Analyte Preparation
Mass Spectrometry
Data Analysis / Interpretation
6
Most Proteomics Experiments Further Purify the
Analyte Using High Performance Liquid
Chromatography (HPLC)
Organic Solvent
H2O
HPLC pumps
7
Two Common Types of Peptide Ionization
Matrix-Assisted Laser Desorption Ionization
(MALDI)
Electrospray Ionization
John Fenn (2002 Nobel Prize)
Koichi Tanaka (2002 Nobel Prize)
8
Perhaps the Most Simple Concept for Mass
Measurements in Mass Spectrometer
Remember there are relationships between mass and
Energy E mc2 or Ek ½ mv2
Time of Flight Mass Spectrometry
Electric field gives ions defined kinetic energy.
Image from Kore Technology Limited
Ekinetic ½ mv2
9
Peptides (Bottom Up Proteomics) Versus Protein
(Top Down)
Tryptic Digest MFSCFLQAGNPQGSRSGFGHNVELVRHASIWVTYH
SEEKLLIPYSDEL MFSCFLQAGNPQGSR SGFGHNVELVR HASIWVT
YHSEEK LLIPYSDEL
10
Another Problem Multiple Peptides may have the
same Mass!
11
Basics of Liquid Chromatography (LC) Tandem Mass
Spectrometry (MS/MS) for Peptides
HPLC pumps
Mass Spectrometer
LC
MS2
MS1
Peptide Fragment Ions
Peptide Ions
low
high
12
Knowing your ABCs and your XYZs In your
Daughter or Fragment Ions
13
A Closer Look at MS2 (For Peptide Identification)
-
Collision- Induced Dissociation ( CID )
14
A Closer Look at MS2 (and Phosphorylation )
-
G-F-S-F-P-V-A-T-G-L-M-E-D-D-G-K-P-R
NH3
COOH
About PowerShow.com