Purification and HPLC-MS analysis of a naturally processed HCMV-derived peptide isolated from the HEK-293T/HLA-E+/Ul40+ cell transfectants and presented at the cell surface in the context of HLA-E.
A new method for isolation and characterization of peptides presented in the context of the nonclassical human leukocytes antigen (HLA) class I molecule HLA-E was developed. A combination of different chromatographic steps coupled with electrospray mass spectrometry allowed us to detect the presence of small amounts of a naturally processed human Cytomegalovirus (HCMV)-derived peptide isolated from the HEK-293T/HLA-E+/UL40+ transfected cells of from HELA cell line. The peptide sequence was confirmed by tandem mass spectrometry (MS/MS). This approach provides a versatile and sensitive method for direct identification of MHC class I-binding peptides that might be derive from different pathogen or tumor-associated proteins.
Millo
E
E
Dipartimento di Medicina Sperimentale, Sezione di Biochimica, c/o CEBR, Università degli Studi di Genova, Genova, Italy. enrico.millo@unige.it
Pietra
G
G
Armirotti
A
A
Vacca
P
P
Mingari
M C
MC
Moretta
L
L
Damonte
G
G
D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000737:chemistry; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003587:Cytomegalovirus; D006680:HLA Antigens / Q000737:chemistry / Q000276:immunology*; D006367:HeLa Cells; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry; D014162:Transfection; D000097625:HLA-E Antigens
D006680:HLA Antigens; D015395:Histocompatibility Antigens Class I; D010455:Peptides; D000097625:HLA-E Antigens