title: Bundling of actin filaments by aorta caldesmon is not related to its regulatory function.
abstract: Ca2+-sensitive thin filaments from vascular smooth muscle were disassembled into their constituent proteins, actin, tropomyosin and caldesmon. Caldesmon bound to both actin and to actin-tropomyosin and inhibited actin-tropomyosin activation of skeletal muscle myosin MgATPase. It also promoted the aggregation of actin or actin-tropomyosin into parallel aligned bundles. Quantitative electron microscopy measurements showed that with 1.1 microM actin-tropomyosin, 1.6 +/- 0.5% (n = 3) of the filaments were in bundles. At 0.073 microM, caldesmon inhibited MgATPase activity by 50%, whereas bundling was 3.0 +/- 1.3% (n = 4). At 0.37 microM caldesmon, MgATPase inhibition was 83% while 28.1 +/- 6.9% (n = 4) of filaments were in bundles. Experiments at 4.4 microM in which MgATPase and bundling were measured in the same samples gave similar results. Small bundles of 2-3 filaments showed the most frequent occurrence at 1.1 microM actin. At 4.4 microM actin the most common bundle size was 3-5 filaments, with the occasional occurrence of large bundles consisting of up to 120 filaments. The incidence of bundling was the same in the presence and absence of tropomyosin. Thus caldesmon can induce the formation of actin bundles but this property bears no relationship to its inhibition of MgATPase activity.
authors.forename: C J
authors.initials: CJ
authors.lastname: Moody
authors.forename: S B
authors.initials: SB
authors.lastname: Marston
authors.forename: C W
authors.initials: CW
authors.lastname: Smith
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D001011:Aorta / Q000378:metabolism; D017301:Ca(2+) Mg(2+)-ATPase / Q000037:antagonists & inhibitors; D002148:Calmodulin-Binding Proteins / Q000502:physiology*; D004789:Enzyme Activation; D066298:In Vitro Techniques; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D012756:Sheep; D014335:Tropomyosin / Q000378:metabolism
chemical_list: D000199:Actins; D002148:Calmodulin-Binding Proteins; D014335:Tropomyosin; D017301:Ca(2+) Mg(2+)-ATPase