RESUMEN
Two-dimensional electrophoresis of total cardiac muscle extracts allows the detection of about 200 protein fractions. In preliminary studies the fraction D-10 protein was characterized in terms of relative molecular mass, isoelectric point and quantitative composition as alpha-tropomyosin. The similarity of the protein to human alpha-tropomyosin was confirmed by the results of analysis of the N-terminal sequence of the D-10 protein eluate in a gas-phase sequencer.
Asunto(s)
Miocardio/química , Tropomiosina/química , Secuencia de Aminoácidos , Electroforesis en Gel Bidimensional , Humanos , Punto Isoeléctrico , Datos de Secuencia Molecular , Tropomiosina/genéticaRESUMEN
Two-dimensional electrophoresis performed according to O'Farrell was used to isolate the myosin light chain I from human heart ventricles. This chain was further used to obtain hybridomas able to secrete monoclonal antibodies. This method permits to increase the yield of monoclonal antibody-secreting hydridomas and to spare effort with regard to cultivation and testing of experimental samples.
Asunto(s)
Ventrículos Cardíacos/química , Miosinas/aislamiento & purificación , Anticuerpos Monoclonales/metabolismo , Electroforesis en Gel Bidimensional , Humanos , Hibridomas/metabolismoRESUMEN
A two-dimensional map of human heart left ventricular proteins for 213 polypeptide fractions has been constructed. A quantitative analysis of variability of the fraction position at 60 selected spots with the use of a CVIT computer system revealed a high reproducibility of the material distribution on electrophoregrams. Differences were found in the protein composition of eight heart muscle atrial and ventricular polypeptide fractions. Left ventricular proteins were shown to be represented by six electrophoretical variants. The methodological peculiarities of construction at the two-dimensional map of heart muscle proteins are discussed.
Asunto(s)
Ventrículos Cardíacos/análisis , Mapeo Peptídico , Proteínas/análisis , Electroforesis en Gel Bidimensional , HumanosRESUMEN
Pattern of myosin light chain in human atrium and ventricles was studied using two-dimensional electrophoresis. Minor fraction was found in ventricles and atria of adult man that coincided in molecular weight, isoelectric point and staining specificity with fetal myosin light chain. The 23 kDa and 24 kDa fractions of auricles were not detected in ventricles.
Asunto(s)
Corazón/embriología , Miocardio/metabolismo , Miosinas/metabolismo , Adulto , Electroforesis en Gel Bidimensional , Edad Gestacional , Atrios Cardíacos/análisis , Atrios Cardíacos/embriología , Atrios Cardíacos/metabolismo , Ventrículos Cardíacos/análisis , Ventrículos Cardíacos/embriología , Ventrículos Cardíacos/metabolismo , Humanos , Miosinas/análisisRESUMEN
Human myocardium proteins synthesized in the course of heart muscle development have been analyzed by two-dimensional electrophoresis. The quantitative change was found in representation of the main retractable proteins in the course of the heart muscle formation (the light myosin chains, tropomyosin, etc.). Four polymorphous variants of myocardium proteins were found, one of which is, possibly connected with the defect in heart development.
Asunto(s)
Corazón/embriología , Proteínas Musculares/análisis , Miocardio/análisis , Envejecimiento/metabolismo , Electroforesis en Gel Bidimensional , Embrión de Mamíferos , HumanosRESUMEN
The study of the polypeptide structure of cardiac muscle proteins of the inbred mice 129 Re + dy/dy by two-dimensional electrophoresis has revealed quantitative and qualitative variations in four polypeptides out of 161 analyzed ones. It is supposed that the protein fractions N 14 and O 13 may contribute to the development of cardiac pathology. Gene-dosage dependence is identified for the fraction O 13.