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Biochemistry 412 Biological Mass Spectrometry February 25 th , 2005. 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. - PowerPoint PPT Presentation
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Biochemistry 412
Biological Mass Spectrometry
February 25th, 2005
There are two major types of “soft ionization” methodsof mass spectrometry for use in analyzingsamples of biological origin:
• Matrix-assisted laser desorption
• Electrospray
Matrix-Assisted Laser Desorption Ionization (MALDI) Mass Spectrometry
Mann et al (2001)Annu. Rev. Biochem. 70, 437.
Electrospray Ionization (ESI) Mass Spectrometry
Griffiths et al (2001)Biochem. J. 355, 545.
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
Example of an ESI Mass Spectrum
Griffiths et al (2001)Biochem. J. 355, 545.
Note:mass-charge ratio
Now for some cool applications
of biological mass spectrometry..!!
Single Nucleotide Polymorphism (SNP)Genotyping by MS
Solid-State DNA Genotyping
Note: extension reactiondone in thepresence of one or more ddNTPs
Tang et al (1999)
Tang et al (1999)
Tang et al (1999)
Generation of Distance Constraints for Protein Structure Determination Using a Combination of Protein Cross-Links and
Mass Spectrometry
Young et al (2000)
Young et al (2000)
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Protein Dynamics, Solvent-Accessibility, and Protein-Protein Interactions
Hamuro et al (2002)
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Hydrogen - Deuterium Exchange and Protein Conformational Fluctuations
Hoofnagle et al (2003) Annu. Rev.Biophys. Biomol. Struct. 32, 1.
Hydrogen - Deuterium Exchange: Schematic Experimental Design
Hoofnagle et al (2003) Annu. Rev.Biophys. Biomol. Struct. 32, 1.
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