Leukophysin: an RNA helicase A-related molecule identified in cytotoxic T cell granules and vesicles.
Leukophysin (LKP) is a 28-kDa protein of CTL and U937 monocytic cells that is located in the membrane of high density granules as well as lighter cytoplasmic granules or vesicles. mAbs to KLP were used to clone a full length cDNA clone with an open reading frame coding for a 235-amino acid polypeptide with a molecular mass of 24.3 kDa and two potential transmembrane regions. The nucleotide sequence was highly homologous to the 3' end of human RNA helicase A. Expression of the LKP was confirmed as a reverse transcriptase-PCR product that may be an alternately spliced product of RNA helicase A. The cDNA contained a repetitive motif that was similar to synaptophysin 1, a protein that is important for synaptic vesicle exocytosis. A polyclonal Ab directed against the 17 carboxyl-terminal amino acids of LKP detected the same 28-kDa granule membrane protein as the D545, one of the mAbs used to clone the cDNA. In addition, the D545 mAb reacted strongly with the GST fusion protein of the bacterially expressed LKP cDNA. In confocal immunofluorescence studies, the anti-LKP peptide Ab reacted with granzyme A-negative granules and vesicles in CD8+ CTL lymphocytes from normal and Chediak-Higashi patients. Thus, based on the expression of the C-terminal LKP epitope, vesicular structures an granules have been detected in CTL that are distinct from classical granzyme-containing cytolytic granules.
Abdelhaleem
M M
MM
Manitoba Institute of Cell Biology, University of Manitoba, Winnipeg, Canada.
Hameed
S
S
Klassen
D
D
Greenberg
A H
AH
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003594:Cytoplasmic Granules / Q000201:enzymology*; D053487:DEAD-box RNA Helicases; D018076:DNA, Complementary / Q000302:isolation & purification; D000939:Epitopes / Q000032:analysis; D053804:Granzymes; D006801:Humans; D007425:Intracellular Membranes / Q000201:enzymology*; D015458:Leukemia, T-Cell; D016403:Lymphoma, Large B-Cell, Diffuse; D008565:Membrane Proteins / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics / Q000302:isolation & purification*; D020365:RNA Helicases; D012316:RNA Nucleotidyltransferases / Q000032:analysis / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D012697:Serine Endopeptidases / Q000032:analysis; D013602:T-Lymphocytes, Cytotoxic / Q000201:enzymology*; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D000939:Epitopes; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D012316:RNA Nucleotidyltransferases; D053804:Granzymes; D012697:Serine Endopeptidases; C509212:GZMA protein, human; C098487:DHX9 protein, human; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases