Adult human liver contains intermediate-type proteasomes with different enzymatic properties

Autores: Gohlke Sabrina, Kloß Alexander, Tsokos Michael, Textoris Taube Kathrin, Keller Christin, Kloetzel Peter-Michael, Dahlmann Burkhardt

Resumen

Background: The 20S proteasome is the proteolytic core of the major intracellular protein degradative system, the ubiquitin-proteasome system. Since little is known about proteasomes of human liver, we have investigated the proteasome spectrum in adult human liver. Material and methods: 20S proteasomes were chromatographically purified from adult human liver and from HuH7 cells. They were divided into subpopulations and subtypes and characterized with regard to their proteolytic activities using short fluorogenic oligo- and long poly-peptide substrates. Their subunit composition was studied by immunoblotting. Results: Proteasomes from adult human liver tissue can be separated into three subpopulations (I, II, III), each of which is composed of several subtypes, which total to a spectrum of 14 different subtypes. Two minor subtypes contain only the immuno-subunits β1i and β5i but not their standard counterparts; all others are intermediate subtypes containing β1 and β5 standard- and β1i and β5i immuno-subunits in various compositions. With regard to the proteolytic activities we observed that a decreasing content of subunit β1i in the subtypes goes along with a decreasing ratio of chymotrypsin-like/caspase-like activity, whereas the degradation rate of a 30 mer polypeptide substrate increased with decreasing β1i content. By comparison, 20S proteasomes from HuH7 cells do not contain immuno-subunits but are pure standard proteasomes, which can be separated into three subtypes. Conclusion: These findings suggest that adult human liver contains a spectrum of 14 different 20S proteasome subtypes with different enzymatic properties reflecting most probably an adaptive response of liver cell functions to challenging factors during lifetime.

Palabras clave: Proteasome-subpopulations proteasome-subtypes proteolytic activities HuH7 cells.

2014-07-07   |   266 visitas   |   Evalua este artículo 0 valoraciones

Vol. 13 Núm.4. Julio-Agosto 2014 Pags. 429-438 Ann Hepatol 2014; 13(4)