RESUMO
Proteins of the human heart muscle were studied using modified two-dimensional electrophoresis. After separation, proteins were electroblotted onto Immobilon P membranes and several protein spots were used for microsequencing analysis. In most cases the proteins analyzed have blocked N-terminal amino acids. In order to study the primary structure of these proteins, hydrolysis in situ by trypsin followed by reversed-phase HPLC and microsequencing of the resulting peptides were performed. Four protein were identified in 8 analyzed fractions, specifically myosin light chain 1 (MLCl-V/sB), fatty-acid binding protein (heart isoform), alpha (B)-crystallin and alpha-tropomyosin. Amino acid sequences of two proteins were not found among human amino acid sequences collected in SWISSPROT bank (v. 21).
Assuntos
Expressão Gênica , Miocárdio/metabolismo , Sequência de Aminoácidos , Western Blotting , Eletroforese em Gel Bidimensional , Humanos , Hidrólise , Dados de Sequência Molecular , Análise de SequênciaRESUMO
Proteins from biopsy of human heart muscle (n = 250) were studied. The supplementary fraction was found in a material from an individuum that coincided in molecular mass but differed in pI from the light chain of myosin usually expressed in the ventricular tissue of the heart muscle (LCM-1 v). The two-dimensional electrophoresis and immunoblotting have shown this supplementary fraction to be a rare allele of LCM-1 v.
Assuntos
Miocárdio/metabolismo , Miosinas/genética , Animais , Western Blotting , Eletroforese em Gel Bidimensional , FosforilaçãoRESUMO
Protein composition of human skeletal muscle impaired with lateral amyotropic sclerosis was studied by means of two-dimensional electrophoresis. Some protein fractions were altered. Characteristic property of all the preparations studied proved to be disappearance of three protein fractions of 35 kDa molecular mass and with pI 5.9, 6.0 and 6.1.