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The identification proteins using mass spectrometry
“mass-mapping / peptide mass-fingerprinting”
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
- amino acid
protein
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
- arginine
protein
- lysine
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
- arginine
- lysine
Trypsin
Recognises and cleaves the protein at the carboxylic acid
of arginine and lysine amino acids
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
- arginine
- lysine
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
- arginine
- lysine
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass
Mass spectrometer
Increasing Mass (longer the peptide the heavier its mass)
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass
Mass spectrometer
Increasing Mass (longer the peptide the heavier its mass)
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass
Mass spectrometer
Increasing Mass (longer the peptide the heavier its mass)
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass
Mass spectrometer
Increasing Mass (longer the peptide the heavier its mass)
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass
Mass spectrometer
Increasing Mass (longer the peptide the heavier its mass)
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass
Mass spectrometer
Increasing Mass (longer the peptide the heavier its mass)
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass
Mass spectrometer
Increasing Mass (longer the peptide the heavier its mass)
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass
Mass spectrometer
Increasing Mass (longer the peptide the heavier its mass)
Dr Kevin MillsInstitute of Child Health, UCL, London
Increasing Mass (longer the peptide the heavier its mass)
The identification proteins using mass spectrometry
Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass
Mass spectrometer
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
11001100 12001200 13001300 14001400 15001500 16001600 17001700 18001800 19001900 20002000 21002100 22002200 23002300 24002400 25002500m/z0
100
%
1283.6
1125.61195.5
2172.2
1531.7
1478.8
1430.7
1586.8
1690.9
1840.1
1953.8
2212.1
Increasing peptide length results in increasing mass of the peptide
Dr Kevin MillsInstitute of Child Health, UCL, London
The identification proteins using mass spectrometry
Trypsin fragment
Dr Kevin MillsInstitute of Child Health, UCL, London
15th Sept 99: spot 3 :in gel digestion MIV( A213)
1200 1400 1600 1800 2000 2200 2400 2600 2800 3000 3200 3400m/z0
100
%
k167n09a 2 (0.155) Sb (15,40.00 ); Sb (1,40.00 ); Sm (SG, 2x3.00); Cm (2:16) TOF LD+ 2.79e41643.9
1231.61405.6
2577.21858.9
2188.93123.3
1antitrypsindeficiency
controlplasma
Identification of Proteins by Peptide Mass Mapping
3D Model of 3D Model of 11-antitrypsin-antitrypsin
In-gel Tryptic
digestion
Comparison of Comparison of peptide masses peptide masses
with with in silico in silico digestion indigestion in datadatabase to obtain base to obtain
““match”match”
Dr Kevin MillsInstitute of Child Health, UCL, London
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The content of this presentation is the copyright of Dr Kevin Mills, UCL Institute of Child Health. Visitors must not copy, distribute, or publish any material from this presentation. It is strictly forbidden to alter this presentation in any way. Any other use of this presentation requires formal permission from Dr Mills.
(c) Dr Kevin Mills, UCL Institute of Child Health, London, 2006.
