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Biochemistry 412 February 24 th , 2002, Lecture Biological Mass Spectrometry

Biochemistry 412 February 24 th , 2002, Lecture Biological Mass Spectrometry. 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.

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Biochemistry 412 February 24 th , 2002, Lecture Biological Mass Spectrometry

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  1. Biochemistry 412 February 24th, 2002, Lecture Biological Mass Spectrometry

  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. 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

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

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

  7. Single Nucleotide Polymorphism (SNP) Genotyping by MS

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

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

  10. Tang et al (1999)

  11. Tang et al (1999)

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

  13. Young et al (2000)

  14. Young et al (2000)

  15. Young et al (2000)

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

  17. Hamuro et al (2002)

  18. Hamuro et al (2002)

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

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

  21. Hamuro et al (2002)

  22. Hamuro et al (2002)

  23. Fragment-Based Drug Discovery

  24. Erlanson et al (2000)

  25. Erlanson et al (2000)

  26. Erlanson et al (2000)

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

  28. Erlanson et al (2000)

  29. Pharmacology Using Imaging Mass Spectrometry

  30. Chaurand et al (2002)

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