Partial purification and MALDI-TOF MS analysis of UN1, a tumor antigen membrane glycoprotein.
UN1 is a membrane glycoprotein that is expressed in immature human thymocytes, a subpopulation of peripheral T lymphocytes, the HPB acute lymphoblastic leukemia (ALL) T-cell line and fetal thymus. We previously reported the isolation of a monoclonal antibody (UN1 mAb) recognizing the UN1 protein that was classified as "unclustered" at the 5th and 6th International Workshop and Conference on Human Leukocyte Differentiation Antigens. UN1 was highly expressed in breast cancer tissues and was undetected in non-proliferative lesions and in normal breast tissues, indicating a role for UN1 in the development of a tumorigenic phenotype of breast cancer cells. In this study, we report a partial purification of the UN1 protein from HPB-ALL T cells by anion-exchange chromatography followed by immunoprecipitation with the UN1 mAb and MALDI-TOF MS analysis. This analysis should assist in identifying the amino acid sequence of UN1.
Department of Clinical and Experimental Medicine, University Magna Graecia of Catanzaro, Via T. Campanella 115, 88100 Catanzaro, Italy. annadela@unicz.it
A
A
de Laurentiis
M
M
Caterino
S
S
Orrù
M
M
Ruoppolo
F
F
Tuccillo
M
M
Masullo
I
I
Quinto
G
G
Scala
P
P
Pucci
C
C
Palmieri
P
P
Tassone
F
F
Salvatore
S
S
Venuta
D000951:Antigens, Neoplasm / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D001943:Breast Neoplasms / Q000737:chemistry / Q000378:metabolism; D045744:Cell Line, Tumor; D005260:Female; D005333:Fetus / Q000737:chemistry / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D051917:Leukosialin; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012795:Sialoglycoproteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013950:Thymus Gland / Q000737:chemistry / Q000378:metabolism
D000951:Antigens, Neoplasm; D006023:Glycoproteins; D051917:Leukosialin; D008565:Membrane Proteins; C465014:SPN protein, human; D012795:Sialoglycoproteins