Mass Spectrometry - Faculté de médecine de l'université d'Ottawa

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Proposed MALDI mechanisms: 1. Primary ionization mechanisms i. Multiphoton ionization ii. Energy pooling and multicenter mo<strong>de</strong>ls iii. Excited-state proton transfer (ESPT) iv. Disproportionation reactions v. Desorption of pre-formed ions vi. Thermal ionization 2. Secondary ionization mechanisms i. Gas-phase proton transfer ii. Gas-phase cationization iii. Electron transfer MALDI is likely a combination of many of these 11/21/2005 34
Matrix Assisted Laser Desorption/Ionisation (MALDI) Unlike ES, MALDI forms predominantly singly charged ions e.g. [M+H] + or adducts (sodium [M+Na] + or potassium [M+K] + ) [M+H] + 22 m/z Sodium = 23 amu Potassium = 39 amu [M+Na] + 38 m/z [M+K] + Brancia FL, Trieste, 12/02/2004 11/21/2005 35
Page 1 and 2: Mass Spectrometry D. Figeys The Ott
Page 3 and 4: A mass spectrum: a plot of the rela
Page 5 and 6: Pumping system Sample introduction
Page 7 and 8: Why is a vacuum necessary? The mean
Page 9 and 10: Various ionisation methods �� E
Page 11 and 12: Electrospray ionization 11/21/2005
Page 13 and 14: Droplet shrinks due to solvent evap
Page 15 and 16: Two examples of common ESI oxidatio
Page 17 and 18: The principal outcome of the electr
Page 19 and 20: ESI + + + + + Direction of electric
Page 21 and 22: ES spectrum of Rho protein 100 % 68
Page 23 and 24: 100 % ES spectrum after deconvoluti
Page 25 and 26: 22+ 11/21/2005 25 22+
Page 27 and 28: Matrix assisted laser desorption io
Page 29 and 30: A standard 100 well UV MALDI sample
Page 31 and 32: Sample plate 2-3 kV + + + + + + + +
Page 33: Why is the matrix so important? �
Page 37 and 38: Pumping system Sample introduction
Page 39 and 40: -(U + V o cosΩt) r r o + − −
Page 41 and 42: http://elchem.kaist.ac.kr/vt/chem-e
Page 43 and 44: What if we place three quadrupoles
Page 45 and 46: Product ion scan or MS/MS MS/MS gas
Page 47 and 48: MS Neutral loss scan gas Neutral lo
Page 49 and 50: Time-of Time of-flight flight (ToF)
Page 51 and 52: Reflectron ion optics: s3 s2 s1 700
Page 53 and 54: Time-Of Time Of-Flight Flight MS 11
Page 55 and 56: Ion traps: Hyperbolic 11/21/2005 55
Page 57 and 58: Ion Trap. Axial Radial toroidal rin
Page 59 and 60: Once in the trap 11/21/2005 59
Page 61 and 62: Ion traps: Linear 11/21/2005 61
Page 63 and 64: Ion traps: Orbital 11/21/2005 63
Page 65 and 66: Lord of the ion rings: Forging of t
Page 67 and 68: LTQ-Orbitrap: LTQ Orbitrap: 2 nd ge
Page 69 and 70: 700 L/s 250 L/s Ion path Q 760 Torr
Page 71 and 72: 11/21/2005 71
Page 73 and 74: GR q0 SL Pusher Q1 Puller q2 TOF Li
Page 75 and 76: Fourier transformed ion cyclotron 1
Page 77 and 78: Protein identification by mass spec
Page 79 and 80: Protein identification: Bioinformat
Page 81 and 82: MALDI-TOF spectrum 11/21/2005 81
Page 83 and 84: Proteomic applications of mass spec
Page 85 and 86:
Protein identification by mass spec
Page 87 and 88:
MEF2A MGRKKIQITRIMDERNRQVTFTKRKFGLM
Page 89 and 90:
Database Searching WWW Sites: Matri
Page 91 and 92:
AGAATVKENSSTRVKHCAAGAAWKAGGARERAAKT
Page 93 and 94:
Select: TNSDIVETIR m n R m 1 IR m 2
Page 95 and 96:
The peptide fragments give us seque
Page 97 and 98:
The 9 step strategy for manual inte
Page 99 and 100:
The 9 step strategy for manual inte
Page 101 and 102:
400 300 200 100 0 Step 2: b 2 and a
Page 103 and 104:
400 300 200 100 0 Step 4: Find y n-
Page 105 and 106:
400 300 200 100 0 Step 6: Find b-io
Page 107 and 108:
400 300 200 100 0 Peptide sequence:
Page 109 and 110:
Relative Abundance 100 80 60 40 20
Page 111 and 112:
Relative Intensity TOF Binary data
Page 113 and 114:
80 40 0 Identification of Proteins
Page 115 and 116:
A) B) C) ∆p Sample introduction:
Page 117 and 118:
A) B) C) HPLC/ autosampler Sample i
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Mass Spectrometry - Faculté de médecine de l'université d'Ottawa

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