Identification of the microvillar 110-kDa calmodulin complex (myosin-1) in kidney.
The epithelial layer lining the proximal convoluted tubule of mammalian kidney contains a brush border of numerous microvilli. These microvilli appear in structure to be very similar to the microvilli on epithelial cells of the small intestine. Microvilli found in both the small intestine and the proximal convoluted tubules in kidney have a core bundle of actin filaments bundled by the accessory proteins villin and fimbrin. Along the length of intestinal microvilli, lateral links can be observed to connect the core bundle of actin filaments to the membrane. These cross-bridges are comprised of a 110-kDa calmodulin complex which belongs to a class of single-headed myosin molecules, collectively referred to as myosin-1. We now report that an analogous calmodulin-binding polypeptide of 105 kDa has been identified in rat kidney cortex. The 105-kDa polypeptide is preferentially found in purified kidney brush borders, can be extracted with ATP, and co-elutes with calmodulin on gel filtration and anion exchange chromatography. Fractions containing the 105-kDa polypeptide exhibit a modest ATPase activity in buffer containing CaCl2. The partially purified 105-kDa polypeptide will bind iodinated calmodulin and will sediment with F-actin in buffer containing ethylene glycol-bis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) or Ca2+. The addition of ATP partially reverses this association with F-actin. These results indicate that myosin-1, in addition to its presence in intestinal brush borders, is present in the brush border of kidney. We also provide preliminary evidence to indicate that the 105-kDa polypeptide is not restricted to tissues possessing a brush border.
Coluccio
L M
LM
Department of Biochemistry, Emory University School of Medicine, Atlanta, GA 30322.
D000818:Animals; D002147:Calmodulin / Q000032:analysis*; D002148:Calmodulin-Binding Proteins; D007672:Kidney Cortex / Q000737:chemistry* / Q000648:ultrastructure; D008871:Microvilli / Q000737:chemistry* / Q000648:ultrastructure; D009218:Myosins / Q000032:analysis* / Q000302:isolation & purification; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D051381:Rats; D014018:Tissue Distribution
C489240:105 kDa calmodulin-binding polypeptide, rat; D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D011506:Proteins; D009218:Myosins