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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.

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Biochemistry 412

Biological Mass Spectrometry

February 25th, 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


Matrix-Assisted Laser Desorption Ionization (MALDI) Mass Spectrometry

Mann et al (2001)

Annu. Rev. Biochem. 70, 437.


Electrospray Ionization (ESI) Mass Spectrometry Spectrometry

Griffiths et al (2001)

Biochem. J.355, 545.


Attractive Features of Mass Spectrometry 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 Spectrometry

Griffiths et al (2001)

Biochem. J.355, 545.

Note:

mass-charge ratio


Now for some cool applications Spectrometry

of biological mass spectrometry..!!


Single Nucleotide Polymorphism (SNP) Spectrometry

Genotyping by MS


Solid-State DNA Genotyping Spectrometry

Note: extension reaction

done in thepresence of

one or more ddNTPs

Tang et al (1999)


Tang et al (1999) Spectrometry


Tang et al (1999) Spectrometry


Generation of Distance Constraints for Spectrometry

Protein Structure Determination Using a

Combination of Protein Cross-Links and

Mass Spectrometry


Young et al (2000) Spectrometry


Young et al (2000) Spectrometry


Young et al (2000) Spectrometry


Hydrogen-Deuterium Exchange and SpectrometryMass Spectrometry to Probe

Protein Dynamics, Solvent-Accessibility,

and Protein-Protein Interactions


Hamuro et al (2002) Spectrometry


Hamuro et al (2002) Spectrometry


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.




Fragment-Based Design

Drug Discovery





The Monophore Binds in the Same Relative Orientation, Regardless of Attachment Site

Erlanson et al (2000)


Erlanson et al (2000) Regardless of Attachment Site


Pharmacology Using Regardless of Attachment Site

Imaging Mass Spectrometry


Chaurand et al (2002) Regardless of Attachment Site


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