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Mouse Flower‐like (mFl), a membrane protein involved in endocytosis?
Author(s) -
MannebachGötz Stefanie,
Frohnweiler Katja,
Wissenbach Ulrich,
Flockerzi Veit
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.1173.13
Subject(s) - biology , fusion protein , microbiology and biotechnology , endocytosis , transgene , gene , membrane protein , receptor , genetics , recombinant dna , membrane
Recently, a novel membrane protein in Drosophila melanogaster was shown to be localized in presynaptic vesicles. It appears to mediate a Ca influx after vesicle fusion with the plasma membrane. Disruption of the corresponding gene leads to endocytic defects in flies. Apparently, this protein could serve as a Ca channel required for endocytosis. In addition it may determine clonal selection by forcing cells to undergo apoptosis. We have identified a protein in mouse which shares 26,3% amino acid sequence identity with the fly protein and which we call mouse Flower‐like or mFl. The mFl protein is encoded by a gene on chromosome 2. In addition to the “full length” mFl protein (mFl A ), 3 shorter variants were identified (mFl B, D, E ). After expression of the His‐SUMO‐mFl A ‐fusion protein in E. coli and removal of the tag the purified protein was used for immunization of rabbits. The anti‐mFl antibody recognizes mFl variants in brain, pancreas, kidney, thymus, liver, lung and colon. Using RT‐PCR mFl A transcripts could be identified additionally in heart, spleen and mast cells. To analyze mFl functions we are generating mFl deficient mouse lines. The targeting strategy for conditional inactivation of the gene offers the option to study the cellular localization of mFl. Currently we have 11 heterozygous mFl transgenic mice; they will be crossed with Cre or Flp expressing mice to get mFl deficient animals.