1. Biological Mass Spectrometry
Biochemistry 412
Biological Mass Spectrometry
February 25th, 2005

2. There are two major types of “soft ionization” methods
of mass spectrometry for use in analyzing
samples of biological origin:
• Matrix-assisted laser desorption
• Electrospray

3. Matrix-Assisted Laser Desorption Ionization (MALDI) Mass Spectrometry
Mann et al (2001)
Annu. Rev. Biochem. 70, 437.

4. Electrospray Ionization (ESI) Mass Spectrometry
Griffiths et al (2001)
Biochem. J. 355, 545.

5. Attractive Features of Mass Spectrometry
• High resolution (can measure mass differences of ~0.1 Da)
• High sensitivity (10-13 to 10-19[!] moles)
• Ability to work with complex samples (MALDI-MS)
• Ability to interface with other separation techniques (e.g., LC)
• “Reasonably” affordable

6. Example of an ESI Mass Spectrum
Griffiths et al (2001)
Biochem. J. 355, 545.
Note:
mass-charge ratio

7. Now for some cool applications of biological mass spectrometry..!!

8. Single Nucleotide Polymorphism (SNP)
Genotyping by MS

10. Note: extension reaction
Solid-State DNA Genotyping
Note: extension reaction
done in thepresence of
one or more ddNTPs
Tang et al (1999)

11. Tang et al (1999)

12. Tang et al (1999)

13. Generation of Distance Constraints for
Protein Structure Determination Using a
Combination of Protein Cross-Links and
Mass Spectrometry

15. Young et al (2000)

16. Young et al (2000)

17. Young et al (2000)

18. Hydrogen-Deuterium Exchange and Mass Spectrometry to Probe
Protein Dynamics, Solvent-Accessibility,
and Protein-Protein Interactions

20. Hamuro et al (2002)

21. Hamuro et al (2002)

22. Hydrogen - Deuterium Exchange and Protein Conformational Fluctuations
Hoofnagle et al (2003) Annu. Rev.
Biophys. Biomol. Struct. 32, 1.

23. Hydrogen - Deuterium Exchange: Schematic Experimental Design
Hoofnagle et al (2003) Annu. Rev.
Biophys. Biomol. Struct. 32, 1.

24. Hamuro et al (2002)

25. Hamuro et al (2002)

26. Fragment-Based
Drug Discovery

28. Erlanson et al (2000)

29. Erlanson et al (2000)

30. Erlanson et al (2000)

31. The Monophore Binds in the Same Relative Orientation, Regardless of Attachment Site
Erlanson et al (2000)

32. Erlanson et al (2000)

33. Imaging Mass Spectrometry
Pharmacology Using
Imaging Mass Spectrometry

35. Chaurand et al (2002)