Proteomic study of human glioblastoma multiforme tissue employing complementary two-dimensional liquid chromatography- and mass spectrometry-based approaches.
An extensive data set comprising 2660 unique protein identifications was obtained for the proteome of a human brain tumor (glioblastoma multiforme) by combining the results of two complementary analytical strategies based on two-dimensional chromatography and mass spectrometry. A bottom-up method, performing peptide separation in both chromatographic dimensions was employed as well as a semi-top-down method, in which intact proteins were separated in the first and tryptic peptides in the second dimension. The identified proteins were assigned to their molecular functions and compared to previously identified proteins of glioblastoma multiforme (= astrocytoma WHO grade IV), lower WHO grade astrocytomas (grade II and III), and nontumor brain tissue. With the use of a subset of 104 identified membrane proteins, the properties of intact protein fractionation in the first dimension of the semi-top-down approach were elucidated in detail. The benefit of the semi-top-down approach was further demonstrated by the identification of a set of endogenous glioblastoma multiforme expressed proteins. These proteins correspond to recombinant antigens which were recently found to be reactive against autoantibodies in glioblastoma multiforme patients. The results indicate the usefulness of the semi-top-down approach for the investigation of immunogenic antigens in human tumor tissue samples.
Melchior
Katja
K
Department of Chemistry, Instrumental Analysis and Bioanalysis, Saarland University, 66123 Saarbrücken, Germany.
Tholey
Andreas
A
Heisel
Sabrina
S
Keller
Andreas
A
Lenhof
Hans-Peter
HP
Meese
Eckart
E
Huber
Christian G
CG
D000951:Antigens, Neoplasm; D001923:Brain Chemistry; D002853:Chromatography, Liquid / Q000379:methods*; D005909:Glioblastoma / Q000378:metabolism*; D006801:Humans; D007526:Isoelectric Point; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008565:Membrane Proteins / Q000737:chemistry; D008875:Middle Aged; D008970:Molecular Weight; D010455:Peptides / Q000032:analysis; D011506:Proteins / Q000032:analysis; D020543:Proteome / Q000032:analysis*; D015203:Reproducibility of Results
D000951:Antigens, Neoplasm; D008565:Membrane Proteins; D010455:Peptides; D011506:Proteins; D020543:Proteome