The 110,000-dalton actin- and calmodulin-binding protein from intestinal brush border is a myosin-like ATPase.
A 110-kDa protein present in chicken intestinal brush-border microvilli is believed to laterally link the actin filament bundle that forms the structural core of the microvilli with the microvillar plasma membrane. We have purified a 110-kDa protein to greater than 95% homogeneity by extraction of brush borders with solution containing 0.6 M KCl and 5 mM ATP, followed by gel filtration chromatography, sedimentation as a complex with exogenous actin, and hydroxylapatite chromatography. The 110-kDa protein-calmodulin complex bound F-actin in the absence but not the presence of ATP and had K+,EDTA-ATPase (0.2 mumol/min/mg) and Ca2+-ATPase (0.2 mumol/min/mg) activities and Mg2+-ATPase activity (0.03 mumol/min/mg) that was not activated by F-actin. The actin-binding and ATPase activities of the complex were similar to those of purified brush-border myosin. However, immunoblot analysis showed no reactivity between the 110-kDa protein and polyclonal antibody against purified chicken brush-border myosin. Also, peptide maps of 110-kDa protein and myosin obtained by limited proteolysis with chymotrypsin and Staphylococcus aureus V8 protease had few, if any, peptides in common. Immunoblot analysis also showed that myosin heavy chain was stable under the conditions of the preparation.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D002148:Calmodulin-Binding Proteins; D002352:Carrier Proteins / Q000378:metabolism*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D018260:Gelsolin; D007163:Immunosorbent Techniques; D007422:Intestines / Q000648:ultrastructure*; D008840:Microfilament Proteins; D008871:Microvilli / Q000201:enzymology; D009218:Myosins / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D011817:Rabbits
D002148:Calmodulin-Binding Proteins; D002352:Carrier Proteins; D018260:Gelsolin; D008840:Microfilament Proteins; C016953:brevin; D010749:Phosphoprotein Phosphatases; D000251:Adenosine Triphosphatases; D009218:Myosins