Thioredoxin expression and localization in human cell lines: detection of full-length and truncated species.
Thioredoxin (Trx) is an intracellular multifunctional 12-kDa protein with a reduction/oxidation (redox) active disulfide constitutively expressed by most cells of the human body. Trx can also be released by cells such as lymphocytes upon activation or oxidative stress exposure and exert a cocytokine and cytoprotective activity. In addition, a truncated 10-kDa form of Trx has been reported. In order to better understand the function of full-length and truncated Trx, we have produced, for the first time, specific monoclonal antibodies, which can discriminate between the two forms. Using these novel antibodies, designated alpha Trx1 to alpha Trx4, a panel of cell lines derived from human B and T lymphocytes, monocytes, granulocytes, and melanomas was analyzed by immunochemical techniques. The cellular distribution differed between the two forms. All lines contained full-length Trx, also located to a minor extent on the cell surface. One exception was the melanoma cell line FM28.4, which did not show any Trx expression. Truncated Trx was present in most cells in minimal amounts only, whereas the monocytic cell lines THP-1 and U-937 expressed high amounts on the cell surface, as shown by flow cytometric analysis of living cells and confocal laser-scanning microscopy. The biological importance and function of the short versus long forms of Trx as detected by the antibodies are discussed.
Sahaf
B
B
Department of Biomedicine and Surgery, Linköping University, Sweden.
Söderberg
A
A
Spyrou
G
G
Barral
A M
AM
Pekkari
K
K
Holmgren
A
A
Rosén
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000954:Antigens, Surface / Q000032:analysis / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D002462:Cell Membrane / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression / Q000502:physiology; D006801:Humans; D007425:Intracellular Membranes / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013879:Thioredoxins / Q000737:chemistry* / Q000235:genetics* / Q000276:immunology; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000502:physiology
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D013879:Thioredoxins