Isolation and characterization of bovine thymus multicatalytic proteinase complex.
The multicatalytic proteinase complex (MPC or proteasome) from bovine thymus was isolated and purified to homogeneity applying a protocol utilizing ion exchange and gel permeation chromatography as major purification tools. The purified complex shows molecular properties that are common for proteasomal molecules (high molecular mass, multisubunit organization, and multiple proteolytic activities) even though a peculiar subunit composition and the presence of specific regulatory mechanisms affecting the assembled proteolytic activities suggest a specialized function for this complex. Thymus proteasome is characterized by the presence of LMP2, LMP7, and LMP10 (MECL1) subunits, which replace the X, Y, and Z subunits. Since a similar complex was previously isolated in bovine spleen, it appears that the proteasomal population containing the LMP subunits is characteristic for organs involved in immune response. Both the thymus and spleen proteasomes are characterized by a marked efficiency in cleaving peptide bonds after branched-chain and aromatic amino acids, indicating that this proteasomal population is most likely involved in intracellular processing of class I antigenic peptides and is an example of an "in vivo" functioning immunoproteasome. However, in spite of several similarities, the complexes isolated from the two lymphoid organs do not show superimposable functional properties, which suggests the presence of organ-specific regulatory mechanisms affecting each of the proteolytic components assembled in the complex.
Eleuteri
A M
AM
Post-Graduate School in Clinical Biochemistry, University of Camerino, Camerino (MC), 62032, Italy. a.eleuteri@cambio.unicam.it
Angeletti
M
M
Lupidi
G
G
Tacconi
R
R
Bini
L
L
Fioretti
E
E
D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D003374:Coumarins / Q000737:chemistry; D003546:Cysteine Endopeptidases / Q000737:chemistry*; D015853:Cysteine Proteinase Inhibitors / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D049934:Isocoumarins; D009097:Multienzyme Complexes / Q000737:chemistry*; D046988:Proteasome Endopeptidase Complex; D013154:Spleen / Q000737:chemistry; D013950:Thymus Gland / Q000737:chemistry*
D003374:Coumarins; D015853:Cysteine Proteinase Inhibitors; D049934:Isocoumarins; D009097:Multienzyme Complexes; C046235:3,4-dichloroisocoumarin; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex