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Glycolytic Enzyme Interactions with the Caveolin Scaffolding Domain (CSD) in a Lymphocyte Model
Author(s) -
Fischer Caitlin N.,
Hardin Christopher D.
Publication year - 2008
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.22.1_supplement.1180.3
Subject(s) - colocalization , transfection , caveolin 3 , glycolysis , microbiology and biotechnology , biology , aldolase a , phosphofructokinase , enzyme , glyceraldehyde 3 phosphate dehydrogenase , caveolin , caveolae , chemistry , biochemistry , dehydrogenase , signal transduction , gene
Previously, we found caveolin (CAV‐1) expressed by transfection in lymphocytes induced caveolae formation and targeted glycolytic enzymes such as aldolase, phosphofructokinase and glyceraldehyde‐3‐phosphate dehydrogenase to the plasma membrane in the CAV‐1 transfected lymphocytes. We hypothesized that if CAV‐1 with a mutant caveolin scaffolding domain (CSD) is expressed in lymphocytes then colocalization of the glycolytic enzymes with CAV‐1 will be reduced. We tested this hypothesis by comparing the colocalization of CAV‐1 with glycolytic enzymes in lymphocytes which expressed either a wild type CAV‐1 (WT) or a mutant CAV‐1 which had either one mutation (SM) or two mutations (DM) in the CSD. Colocalization analysis by confocal microscopy for CAV‐1 and ALD was 0.77 in CAV‐1 WT lymphocytes, 0.24 in CAV‐1 SM lymphocytes, and 0.59 in CAV‐1 DM lymphocytes. Analysis of colocalization of the enzymes PFK, GAPDH, and ALD with CAV‐1 averaged 0.65 for CAV‐1 WT cells, 0.49 for CAV‐1 SM cells and 0.51 for the CAV‐1 DM cells. The shift in distribution of glycolytic enzymes and CAV‐1 in the CAV‐1 WT, the CAV‐1 SM or DM CAV‐1 types indicates that a single mutation to the CSD reduces glycolytic enzyme membrane targeting, and two mutations in the CSD produces CAV‐1 retention in the cytoplasm, both suggesting that an intact CSD domain is essential for CAV‐1 targeting of glycolytic enzymes to the membrane. Support NIH DK60668.