Fragmentation of multiply-charged intact protein ions using MALDI TOF-TOF mass spectrometry.
Top down proteomics in a TOF-TOF instrument was further explored by examining the fragmentation of multiply charged precursors ions generated by matrix-assisted laser desorption ionization. Evaluation of sample preparation conditions allowed selection of solvent/matrix conditions and sample deposition methods to produce sufficiently abundant doubly and triply charged precursor ions for subsequent CID experiments. As previously reported, preferential cleavage was observed at sites C-terminal to acidic residues and N-terminal to proline residues for all ions examined. An increase in nonpreferential fragmentation as well as additional low mass product ions was observed in the spectra from multiply charged precursor ions providing increased sequence coverage. This enhanced fragmentation from multiply charged precursor ions became increasingly important with increasing protein molecular weight and facilitates protein identification using database searching algorithms. The useable mass range for MALDI TOF-TOF analysis of intact proteins has been expanded to 18.2 kDa using this approach.
Liu
Zhaoyang
Z
Department of Cell and Molecular Pharmacology, Medical University of South Carolina, Charleston, South Carolina 29425, USA.
Schey
Kevin L
KL
D000595:Amino Acid Sequence; D000818:Animals; D001363:Awards and Prizes; D002417:Cattle; D029968:Escherichia coli Proteins / Q000032:analysis / Q000737:chemistry; D006736:Horses; D007782:Lactoglobulins / Q000032:analysis / Q000737:chemistry; D008894:Milk Proteins / Q000032:analysis / Q000737:chemistry; D008969:Molecular Sequence Data; D009211:Myoglobin / Q000032:analysis / Q000737:chemistry; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013879:Thioredoxins / Q000032:analysis / Q000737:chemistry; D025801:Ubiquitin / Q000032:analysis / Q000737:chemistry
D029968:Escherichia coli Proteins; D007782:Lactoglobulins; D008894:Milk Proteins; D009211:Myoglobin; D011506:Proteins; D025801:Ubiquitin; D013879:Thioredoxins
J Am Soc Mass Spectrom. 2002 Jul;13(7):784-91
12148803
J Am Soc Mass Spectrom. 2005 Apr;16(4):482-90
15792717
Anal Chem. 1995 Nov 1;67(21):3990-9
8633762
J Am Soc Mass Spectrom. 1995 Dec;6(12):1183-9
24214069
Anal Chem. 2002 Feb 1;74(3):577-83
11838679
Rapid Commun Mass Spectrom. 2003;17(20):2343-8
14558136
Anal Chem. 2001 Jul 15;73(14):3274-81
11476225
Anal Chem. 1993 Nov 1;65(21):3015-23
8256865
Anal Chem. 1996 Mar 1;68(5):859-66
21619182
Mass Spectrom Rev. 2005 Nov-Dec;24(6):931-58
15706594
Anal Chem. 1996 Jan 1;68(1):31-7
8779435
Proc Natl Acad Sci U S A. 2002 Dec 10;99(25):15863-8
12444260
Anal Chem. 1994 Sep 15;66(18):2809-15
7526742
J Am Soc Mass Spectrom. 2001 May;12(5):565-70
11349954
Anal Chem. 2006 Feb 15;78(4):1218-27
16478115
Anal Chem. 2005 Nov 15;77(22):7455-61
16285700
J Am Chem Soc. 2005 Sep 14;127(36):12627-39
16144411
Anal Chem. 1994 Sep 15;66(18):2801-8
7978294
Anal Chem. 2000 Feb 1;72(3):563-73
10695143
Science. 1990 Apr 13;248(4952):201-4
2326633
J Am Chem Soc. 2002 Jun 5;124(22):6407-20
12033872
Curr Opin Biotechnol. 2002 Feb;13(1):57-64
11849959
J Mass Spectrom. 2000 Jan;35(1):1-12
10633229
J Am Soc Mass Spectrom. 2001 Jul;12(7):873-6
11444611
J Am Chem Soc. 2002 Jan 30;124(4):672-8
11804498
Anal Chem. 2001 Jan 1;73(1):19-22
11195502
Anal Chem. 1998 Oct 15;70(20):4410-6
9796424
J Mass Spectrom. 2003 Mar;38(3):245-56
12644985
Proc Natl Acad Sci U S A. 2005 Jul 5;102(27):9463-8
15983376
J Proteome Res. 2005 Jul-Aug;4(4):1403-12
16083293
Anal Chem. 2000 Mar 1;72(5):899-907
10739190
Angew Chem Int Ed Engl. 2003 Oct 20;42(40):4900-4
14579433
Rapid Commun Mass Spectrom. 2003;17(16):1809-14
12876680
J Am Soc Mass Spectrom. 2001 Mar;12(3):245-9
11281599
J Am Soc Mass Spectrom. 2006 Jul;17(7):923-31
16698278