Premium
Nitric oxide synthase and dynamin interactions in the renal inner medulla
Author(s) -
Hyndman Kelly Anne,
Musall Jacqueline B,
Xue Jing,
Pollock Jennifer
Publication year - 2009
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.23.1_supplement.602.6
Subject(s) - dynamin , endocytosis , nos1 , microbiology and biotechnology , nitric oxide , chemistry , gene isoform , nitric oxide synthase , hek 293 cells , biology , medicine , endocrinology , biochemistry , receptor , gene
The movements of proteins in and out of the plasma membrane are important steps in regulating protein function. Dynamin (DNM) is a large GTPase that forms a helical collar around invaginated vesicles, and "pinches" it from the membrane during endocytosis. Recently, nitric oxide (NO) was reported to enhance DNM‐mediated endocyotosis through s‐nitrosylation of DNM in HEK cells, thus suggesting an important regulatory link between NO and endocytosis. We hypothesize that NO/DNM interactions are important mediators of ion channel function in the kidney. The purpose of this study was to determine which NOS and DNM isoforms directly interact in the renal inner medulla (IM). Sprague Dawley rats on a normal sodium diet were anesthetized and the IMs dissected and pooled per animal. These samples were immunoprecipitated with anti‐DNM and immunoblotted with NOS isoform specific antibodies. We determined there are direct interactions between DNM/NOS1 (alpha and beta splice variants), and DNM/NOS3 in vivo . In addition, DNMs1‐3 and NOS1 and NOS3 were immunolocalized to the IM collecting ducts (IMCD). We further confirmed our in vivo results by cotransfecting mIMCD‐3 and COS 7 cells with NOS1 and dynamin‐GFP constructs and determined these two proteins do directly interact in vitro . We conclude that DNM and NOS directly interact in the IMCD, leading us to further hypothesize that NO regulation of DNM is an important mechanism in regulating ion channel density in the IMCD.