Biochemistry 412
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Biochemistry 412 Biological Mass Spectrometry February 25 th , 2005 PowerPoint PPT Presentation


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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 25 th , 2005

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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

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 reaction

done 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)


Young et al (2000)


Hydrogen-Deuterium Exchange and Mass Spectrometry to Probe

Protein Dynamics, Solvent-Accessibility,

and Protein-Protein Interactions


Hamuro et al (2002)


Hamuro et al (2002)


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.


Hamuro et al (2002)


Hamuro et al (2002)


Fragment-Based

Drug Discovery


Erlanson et al (2000)


Erlanson et al (2000)


Erlanson et al (2000)


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

Erlanson et al (2000)


Erlanson et al (2000)


Pharmacology Using

Imaging Mass Spectrometry


Chaurand et al (2002)


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