scholarly journals Extraction Protocols for Individual Zebrafish's Ventricle Myosin and Skeletal Muscle Actin for In vitro Motility Assays

2017 ◽  
Vol 8 ◽  
Author(s):  
Lisa-Mareike Scheid ◽  
Cornelia Weber ◽  
Nasrin Bopp ◽  
Matias Mosqueira ◽  
Rainer H. A. Fink
Keyword(s):  
Skeletal Muscle ◽  
Muscle Actin ◽  
In Vitro Motility

scholarly journals Cyclophosphamide treatment evoked side effect on skeletal muscle actin, monitored by DSC

2021 ◽  
Author(s):  
Péter Farkas ◽  
Dávid Szatmári ◽  
Franciska Könczöl ◽  
Dénes Lőrinczy
Keyword(s):  
Skeletal Muscle ◽  
Side Effect ◽  
Main Peak ◽  
Muscle Actin ◽  
Scanning Calorimetry ◽  
Drug Induced ◽  
Cyclophosphamide Treatment ◽  
Intact Muscle ◽  
Binding Cleft

AbstractSeveral kind of drugs—used in cancer treatments—such as cyclophosphamide (CP) can also trigger a disease classified as toxic polyneuropathy. Polyneuropathy is a simultaneous malfunction of several peripheral nerves, typical side effect of a cancer therapy. In our previous study, we used CP treated in vitro animal model (Guinea pig) with a comparable dosage and time handling of human protocol to show evidences of this drug-induced effects. We could show a dose-dependent difference between in Tm and ΔHcal of untreated and treated samples assigned to their intact muscle and nerve, blood plasma and red blood cells. In our current study we analyze this side effect on skeletal muscle actin (prepared from m. psoas of rabbit) by DSC (differential scanning calorimetry), to follow the possible consequence of drug treatment on the “activator” of muscle contraction. We have demonstrated that run of DSC curves, Tms together with the ΔHcal exhibit clear CP effect. In case of Ca2+ G actin it is manifested in a well separated second high denaturing temperature as a consequence of CP binding into the cleft. This way the nucleotide binding cleft with subdomains 1 and 3 becomes less flexible, indicating clear sensitivity to CP treatment. In F-actin samples, the main peak represents the thermal denaturation of subdomains 1 and 3, and the increased calorimetric enthalpy administrating Ca2+ as well as CP refers to a more rigid structure. These alterations can be the molecular background in the malfunction of muscle in case of polyneuropathy after CP treatment.

In Vitro Motility of Skeletal Muscle Myosin and Its Proteolytic Fragments1

1990 ◽  
Vol 107 (5) ◽  
pp. 671-679 ◽  
Author(s):  
Kingo Takiguchi ◽  
Hiroshi Hayashi ◽  
Eiji Kurimoto ◽  
Sugie Higasshi-Fujime
Keyword(s):  
Skeletal Muscle ◽  
Skeletal Muscle Myosin ◽  
In Vitro Motility ◽  
Muscle Myosin

In vitro motility studies of a DCM mutation in cardiac muscle actin

2007 ◽  
Vol 42 (6) ◽  
pp. S162
Author(s):  
E Dyer ◽  
D Wells ◽  
C Redwood ◽  
S Marston
Keyword(s):  
Cardiac Muscle ◽  
Muscle Actin ◽  
In Vitro Motility

Glutathione reverses early effects of glycation on myosin function

2003 ◽  
Vol 285 (2) ◽  
pp. C419-C424 ◽  
Author(s):  
B. Ramamurthy ◽  
A. Daniel Jones ◽  
L. Larsson
Keyword(s):  
Skeletal Muscle ◽  
Extracellular Proteins ◽  
Dependent Manner ◽  
Skeletal Muscle Function ◽  
In Vitro Motility Assay ◽  
In Vitro Motility ◽  
Glucose Exposure ◽  
And Function ◽  
Dose Dependent

Nonenzymatic glycosylation (glycation) has been recognized as an important posttranslational modification underlying alterations of structure and function of extracellular proteins during aging and diabetes. Intracellular proteins may also be affected by this modification, and glycation has been suggested to contribute to aging-related impairment in skeletal muscle function. Glycation is the chemical reaction of reducing sugars with primary amino groups resulting in the formation of irreversible advanced glycation end products. Glutathione is an abundant tripeptide in skeletal muscle. To understand the effect of glutathione on glycated myosin function, we used a single-fiber in vitro motility assay in which myosin is extracted from a single muscle fiber segment to propel fluorescent-labeled actin filaments. Myosin function responded to glucose exposure in a dose-dependent manner, i.e., motility speeds were reduced by 10, 34, and 90% of preincubation values after 30-min exposure to 1, 3, and 6 mM glucose, respectively. The 30-min 6 mM glucose incubation was followed by a 20-min 10 mM glutathione incubation. Glutathione treatment restored motility (0.98 ± 0.06 μm/s, n = 3; P < 0.001) after glucose exposure (0.10 ± 0.07 μm/s, n = 3), close to preincubation levels (1.12 ± 0.06 μm/s, n = 3). It is concluded that glucose modifies myosin function in a dose-dependent manner and that glutathione reverses the effect of glucose on myosin function.

scholarly journals Acceleration of the sliding movement of actin filaments with the use of a non-motile mutant myosin in in vitro motility assays driven by skeletal muscle heavy meromyosin

PLoS ONE ◽  
2017 ◽  
Vol 12 (7) ◽  
pp. e0181171 ◽  
Author(s):  
Kohei Iwase ◽  
Masateru Tanaka ◽  
Keiko Hirose ◽  
Taro Q. P. Uyeda ◽  
Hajime Honda
Keyword(s):  
Skeletal Muscle ◽  
Actin Filaments ◽  
Heavy Meromyosin ◽  
In Vitro Motility

scholarly journals Muscle actin filaments bind pituitary secretory granules in vitro.

1977 ◽  
Vol 73 (1) ◽  
pp. 78-87 ◽  
Author(s):  
R E Ostlund ◽  
J T Leung ◽  
D M Kipnis
Keyword(s):  
Skeletal Muscle ◽  
Metal Ions ◽  
Secretory Granule ◽  
Actin Filaments ◽  
Anterior Pituitary ◽  
Secretory Granules ◽  
Muscle Actin ◽  
Viscous Solutions ◽  
Styrene Butadiene

Hog anterior pituitary secretory granules sediment at 3,000 g. When rat or rabbit skeletal muscle actin filaments are present with the granules, the sedimentation decreases markedly. Depolymerized actin or viscous solutions of Ficoll and collagen have no effect on granule sedimentation. With this assay, actin filaments bind secretory granules (consisting of the proteinaceous core plus limiting membrane), secretory granule membranes, mitochondria, artificial lecithin liposomes, and styrene-butadiene microspheres, but have little or no interaction with membrane-free secretory granule cores and albumin microspheres. A secretory granule-actin complex sedimentable between 3,000 g and 25,000 g can be isolated. Metal ions, nucleotides, salts, dithiothreitol, or pretreatment of the granules with trypsin do not destroy the binding, which appears to be a lipophilic interaction.

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