prion project - update mass spectrometry of prions andy lo, béla reiz, messele fentabil dr. liang...

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Prion Project - Update Mass Spectrometry of Prions Andy Lo, B é la Reiz , Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

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Page 1: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Prion Project - UpdateMass Spectrometry of Prions

Andy Lo, Béla Reiz, Messele Fentabil

Dr. Liang Li

Department of Chemistry

University of Alberta

Page 2: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Mass Spectrometry (MS)

Mass Spectrometry is an instrumental method used to identify chemical substances by means of the separation of their gaseous ions using magnetic and/or electric fields according to their mass-to-charge ratio.

Page 3: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

MS in PROTEOMICS

Molecular weight of proteins

Masses of peptides in a mixture

Masses of fragment ions of a peptide

Page 4: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Experimental Procedure

Protein

Digestion (Peptides)

LC-MALDIMS

MS

MSMS

MS

Database Search

Page 5: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Protein MS

Page 6: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Experimental Procedure

Protein

Digestion (Peptides)

LC-MALDIMS

MS

MSMS

MS

Database Search

Page 7: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

PrP Tryptic Digest - MS

Page 8: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

PrP&Myo Tryptic Digest - MS

Page 9: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Experimental Procedure

Protein

Digestion (Peptides)

LC-MALDIMS

MS

MSMS

MS

Database Search

Page 10: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

PrP&Myo Tryptic DigestLC-MALDI-MS

Page 11: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Experimental Procedure

Protein

Digestion (Peptides)

LC-MALDIMS

MS

MSMS

MS

Database Search

Page 12: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

PrP&Myo Tryptic DigestLC-MALDI-MSMS

Page 13: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Experimental Procedure

Protein

Digestion (Peptides)

LC-MALDIMS

MS

MSMS

MS

Database Search

Page 14: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Database Search Results

Page 15: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Database Search Results

Page 16: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Database Search Results

Page 17: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Database Search Results

Page 18: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

PrP Chymotryptic DigestLC-MALDI

Page 19: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

PrP Chymotryptic DigestResults

Page 20: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

PrP Chymotryptic DigestResults

Page 21: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

PrP Chymotryptic DigestResults

Page 22: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

PrP Chymotryptic DigestResults

Page 23: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Future Work

Chemical surface modification of surface exposed residues using:

N-bromosuccinimide (NBS) for Trp exposed residues

Hydroxylation of surface exposed Cys, Trp, Tyr, Met

Page 24: Prion Project - Update Mass Spectrometry of Prions Andy Lo, Béla Reiz, Messele Fentabil Dr. Liang Li Department of Chemistry University of Alberta

Thanks!

Dr. Liang Li Andy and Messele All Li Group members