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 25 th 2005

Biochemistry 412

Biological Mass Spectrometry

February 25th, 2005


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


Biochemistry 412 biological mass spectrometry february 25 th 2005

Matrix-Assisted Laser Desorption Ionization (MALDI) Mass Spectrometry

Mann et al (2001)

Annu. Rev. Biochem. 70, 437.


Biochemistry 412 biological mass spectrometry february 25 th 2005

Electrospray Ionization (ESI) Mass Spectrometry

Griffiths et al (2001)

Biochem. J.355, 545.


Biochemistry 412 biological mass spectrometry february 25 th 2005

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


Biochemistry 412 biological mass spectrometry february 25 th 2005

Example of an ESI Mass Spectrum

Griffiths et al (2001)

Biochem. J.355, 545.

Note:

mass-charge ratio


Biochemistry 412 biological mass spectrometry february 25 th 2005

Now for some cool applications

of biological mass spectrometry..!!


Biochemistry 412 biological mass spectrometry february 25 th 2005

Single Nucleotide Polymorphism (SNP)

Genotyping by MS


Biochemistry 412 biological mass spectrometry february 25 th 2005

Solid-State DNA Genotyping

Note: extension reaction

done in thepresence of

one or more ddNTPs

Tang et al (1999)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Tang et al (1999)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Tang et al (1999)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Generation of Distance Constraints for

Protein Structure Determination Using a

Combination of Protein Cross-Links and

Mass Spectrometry


Biochemistry 412 biological mass spectrometry february 25 th 2005

Young et al (2000)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Young et al (2000)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Young et al (2000)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Hydrogen-Deuterium Exchange and Mass Spectrometry to Probe

Protein Dynamics, Solvent-Accessibility,

and Protein-Protein Interactions


Biochemistry 412 biological mass spectrometry february 25 th 2005

Hamuro et al (2002)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Hamuro et al (2002)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Hydrogen - Deuterium Exchange and Protein Conformational Fluctuations

Hoofnagle et al (2003) Annu. Rev.

Biophys. Biomol. Struct.32, 1.


Biochemistry 412 biological mass spectrometry february 25 th 2005

Hydrogen - Deuterium Exchange: Schematic Experimental Design

Hoofnagle et al (2003) Annu. Rev.

Biophys. Biomol. Struct.32, 1.


Biochemistry 412 biological mass spectrometry february 25 th 2005

Hamuro et al (2002)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Hamuro et al (2002)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Fragment-Based

Drug Discovery


Biochemistry 412 biological mass spectrometry february 25 th 2005

Erlanson et al (2000)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Erlanson et al (2000)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Erlanson et al (2000)


Biochemistry 412 biological mass spectrometry february 25 th 2005

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

Erlanson et al (2000)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Erlanson et al (2000)


Biochemistry 412 biological mass spectrometry february 25 th 2005

Pharmacology Using

Imaging Mass Spectrometry


Biochemistry 412 biological mass spectrometry february 25 th 2005

Chaurand et al (2002)


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