Identification of Proteins in Renaissance Paintings by Proteomics.

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Identification of Proteins in Renaissance
Paintings by Proteomics.

• Caroline Tokarski, Elisabeth Martin, Christian
  Rolando, and Cecile Cren-Olive
• Anal Chem. 2006,vol. 78, 1494-1502

2
Background

• Preservation of Paintings.
• A Paintings layers -Preparation coat
  -White Coat
• -Pictorial Coat
• -Varnish
• Pictorial Coat- Paint pigments and
  binder

3
Previous Techniques.

• Protein Detection
• Organic dyes
• immunodetection
• Chromatography
• Thin layer Chromatography
• Gas Chromatography
• Pyrolysis G.C.
• HPLC coupled with UV-Visible Spectroscopy
• Raman Spectroscopy

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Difficulties/Challenges

• To find appropriate extracting conditions of
  proteins from the binding medium without
  hydrolysis in amino acids
• Develop an analytical method closely adapted to
  the small sample amount.

5
Methods

• Paintings
• Benedetto Bonfigli Triptych, The Virgin and
  Child, St. John the Baptist, St. Sebastian.
• Niccolo di Pietro Gerini, The Virgin and Child.
• Paint Media Emulsion.
• Model using whole protein- Ovalbumin (10 w/v)
  and lead white (50w/v).
• Egg Painting Model- Oil emulsion-linseed oil
  (20v/v)
• Sample Extraction and Enzymatic Hydrolysis.
• Sample Analysis and Database search.
• MALDI-TOF DE-STR- Linear Mode and Reflector Mode.
• Nanochromatography/Q-STAR Pulsar Mass
  Spectrometer (ESI-Qq-TOF)MS/MS
• Protein Prospector and MASCOT(NCBI)

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Protein Extraction

• 1 of TFA ground with synthetic resin
• Ultrasonic Baths
• Centrifugation
• MALDI-TOF Mass Spectrometer

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Pre- Check-Maldi-TOF MS Spectrum in Linear Mode
of Ovalbumin
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MALDI-TOF MS spectrum in linear mode of protein
extract from egg based model.
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MALDI-TOF MS spectrum in reflector mode of
hydrolyzed protein extract
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MS/MS spectrum of triply charged ion at m/z
522.28 from hydrolyzed extract of egg-based
painting.
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Application of Developed Method to Renaissance
Paintings

• Benedetto Bonfiglis Triptych
• The Virgin, Child, St. John the Baptist, St.
  Sebastian

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MS/MS spectrum of the triply charged ion at m/z
551.61 from hydrolyzed extract of Bonfiglis
Triptych.
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MS/MS spectrum of the doubly charged ion at m/z
714.82 from the hydrolyzed extract of Bonfigilis
triptych.
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MS/MS spectrum of the monocharged ion at m/z
402.28 from the hydrolyzed extract of Bonfiglis
triptych.
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Niccolo di Pietro Gerini - The Virgin and Child
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MS/MS spectrum of the triply charged ion at m/z
519.23 from the hydrolyzed extract of Gerinis
Painting.
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Conclusion

• Identification of Binder Proteins
• Binder complexity