Abstract

In this project the Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI) concept is being extended for use in Time-of-Flight Static Secondary Ion Mass Spectrometry (ToF-SSIMS). The overall aim of the project is to use MALDI matrix molecules to increase both the ion yields of intact proteins and peptides and to reduce fragmentation caused by Cs+ ion beam impact that occurs in standard SIMS analysis. Initial work is directed towards the detection of intact model peptide molecules. Model peptides with ?-sheet (YGLKLKLKL-NH2 Mw=1075.42 amu) and ?-helix (YG(LKKLLKL)2-NH2 Mw=1911.34 amu)) tertiary structures (synthesized using standard FMOC solid phase synthesis) are being used in conjunction with standard MALDI matrices [2,5-dihydroxybenzoic acid (DHB) and 3,5-dimethoxy-4-hydroxycinnamic (sinapinic acid (SA))] to facilitate generation of molecular ions upon irradiation with the SIMS Cs+ ion >source. SIMS imaging with micrometer spatial resolution of mat rix/peptid es crystals on Si wafers has to date suggested that this mode of analysis can be applied to specifically map the distribution of peptide molecules within MALDI crystals.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001296-17
Application #
6492593
Study Section
Project Start
2001-09-01
Project End
2002-08-31
Budget Start
Budget End
Support Year
17
Fiscal Year
2001
Total Cost
Indirect Cost

  Institution

Name
University of Washington
Department
Type
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195

  Publications

Tyler, Bonnie J; Peterson, Richard E (2013) Dead-time correction for time-of-flight secondary-ion mass spectral images: a critical issue in multivariate image analysis. Surf Interface Anal 45:475-478
Tyler, B J; Bruening, C; Rangaranjan, S et al. (2011) TOF-SIMS imaging of adsorbed proteins on topographically complex surfaces with Bi(3) (+) primary ions. Biointerphases 6:135
Medzihradszky, Katalin F (2008) Characterization of site-specific N-glycosylation. Methods Mol Biol 446:293-316
Medzihradszky, Katalin F (2005) Peptide sequence analysis. Methods Enzymol 402:209-44
Sanders, Joan E; Lamont, Sarah E; Karchin, Ari et al. (2005) Fibro-porous meshes made from polyurethane micro-fibers: effects of surface charge on tissue response. Biomaterials 26:813-8
Medzihradszky, Katalin F (2005) In-solution digestion of proteins for mass spectrometry. Methods Enzymol 405:50-65
Medzihradszky, Katalin F (2005) Characterization of protein N-glycosylation. Methods Enzymol 405:116-38
Cheng, Xuanhong; Wang, Yanbing; Hanein, Yael et al. (2004) Novel cell patterning using microheater-controlled thermoresponsive plasma films. J Biomed Mater Res A 70:159-68
Wagner, Victoria E; Koberstein, Jeffrey T; Bryers, James D (2004) Protein and bacterial fouling characteristics of peptide and antibody decorated surfaces of PEG-poly(acrylic acid) co-polymers. Biomaterials 25:2247-63
Tsai, W B; Shi, Q; Grunkemeier, J M et al. (2004) Platelet adhesion to radiofrequency glow-discharge-deposited fluorocarbon polymers preadsorbed with selectively depleted plasmas show the primary role of fibrinogen. J Biomater Sci Polym Ed 15:817-40

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