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The identification proteins using mass spectrometry

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Presentation on theme: "The identification proteins using mass spectrometry"— Presentation transcript:

1 The identification proteins using mass spectrometry
“mass-mapping / peptide mass-fingerprinting” Dr Kevin Mills Institute of Child Health, UCL, London

2 protein The identification proteins using mass spectrometry
- amino acid Dr Kevin Mills Institute of Child Health, UCL, London

3 protein The identification proteins using mass spectrometry - arginine
- lysine Dr Kevin Mills Institute of Child Health, UCL, London

4 The identification proteins using mass spectrometry
Recognises and cleaves the protein at the carboxylic acid of arginine and lysine amino acids Trypsin - arginine - lysine Dr Kevin Mills Institute of Child Health, UCL, London

5 The identification proteins using mass spectrometry
- arginine - lysine Dr Kevin Mills Institute of Child Health, UCL, London

6 The identification proteins using mass spectrometry
- arginine - lysine Dr Kevin Mills Institute of Child Health, UCL, London

7 The identification proteins using mass spectrometry
Dr Kevin Mills Institute of Child Health, UCL, London

8 The identification proteins using mass spectrometry
Dr Kevin Mills Institute of Child Health, UCL, London

9 The identification proteins using mass spectrometry
Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass Dr Kevin Mills Institute of Child Health, UCL, London

10 The identification proteins using mass spectrometry
Mass spectrometer Increasing Mass  (longer the peptide the heavier its mass) Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass Dr Kevin Mills Institute of Child Health, UCL, London

11 The identification proteins using mass spectrometry
Mass spectrometer Increasing Mass  (longer the peptide the heavier its mass) Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass Dr Kevin Mills Institute of Child Health, UCL, London

12 The identification proteins using mass spectrometry
Mass spectrometer Increasing Mass  (longer the peptide the heavier its mass) Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass Dr Kevin Mills Institute of Child Health, UCL, London

13 The identification proteins using mass spectrometry
Mass spectrometer Increasing Mass  (longer the peptide the heavier its mass) Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass Dr Kevin Mills Institute of Child Health, UCL, London

14 The identification proteins using mass spectrometry
Mass spectrometer Increasing Mass  (longer the peptide the heavier its mass) Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass Dr Kevin Mills Institute of Child Health, UCL, London

15 The identification proteins using mass spectrometry
Mass spectrometer Increasing Mass  (longer the peptide the heavier its mass) Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass Dr Kevin Mills Institute of Child Health, UCL, London

16 The identification proteins using mass spectrometry
Mass spectrometer Increasing Mass  (longer the peptide the heavier its mass) Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass Dr Kevin Mills Institute of Child Health, UCL, London

17 The identification proteins using mass spectrometry
Mass spectrometer Increasing Mass  (longer the peptide the heavier its mass) Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass Dr Kevin Mills Institute of Child Health, UCL, London

18 The identification proteins using mass spectrometry
Mass spectrometer Increasing Mass  (longer the peptide the heavier its mass) Tryptic digestion of the protein provides peptide fragments of varying length and therefore mass Dr Kevin Mills Institute of Child Health, UCL, London

19 The identification proteins using mass spectrometry
Increasing peptide length results in increasing mass of the peptide 100 1283.6 2172.2 1531.7 % 1586.8 1478.8 1690.9 1125.6 2212.1 1195.5 1430.7 1840.1 1953.8 m/z 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000 2100 2200 2300 2400 2500 Dr Kevin Mills Institute of Child Health, UCL, London

20 The identification proteins using mass spectrometry
Trypsin fragment Dr Kevin Mills Institute of Child Health, UCL, London

21 Identification of Proteins by Peptide Mass Mapping
1antitrypsin deficiency Comparison of peptide masses with in silico digestion in data base to obtain “match” control plasma In-gel Tryptic digestion 3D Model of a1-antitrypsin Dr Kevin Mills Institute of Child Health, UCL, London

22 Content on this website is the copyright of the Great Ormond Street Hospital for Children and the Institute of Child Health unless otherwise stated. Where copyright applies, visitors can download material for private research, study or in-house use only. Visitors must not copy, distribute, or publish any material from this website. Any other use of copyright material requires the formal permission of the Great Ormond Street Hospital for Children and the Institute of Child Health. Information specifically for patients and families has been reviewed for accuracy, but we hold no responsibility for alterations made thereafter without our consent. 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.


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