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Lysozyme degradation by the bovine multicatalytic proteinase complex (proteasome): evidence for a nonprocessive mode of degradation.

Abstract：

:The multicatalytic proteinase complex (MPC, proteasome) is composed of 28 subunits organized into four rings surrounding a water-filled canal. The catalytic centers face the inner canal confining protein substrates to an enclosed space. Experimental findings obtained with MPC from archaebacteria suggest that degradation of proteins by the complex is processive and have led to the proposal that the lengths of the peptides formed during degradation depend on the distances between active sites in the catalytic chamber. To test whether these postulates are valid for the MPC from a higher organism, we examined the size distributions of products formed early versus late in the course of protein degradation using reduced carboxamidomethylated lysozyme (RCM-lysozyme) and MPC from bovine spleen and pituitary. The majority of final degradation products ranged in length from 6 to 20 amino acids without a clear predilection for peptides of a particular, uniform size. Our observations suggest that selection of cleavage sites is governed by the amino acid sequence specificity of the MPC catalytic sites rather than the distances between the active sites. Early in the course of degradation, peptides with masses between 5 and 10 kDa accumulated in more than 80-fold molar excess over the MPC, indicating dissociation of large, partially degraded intermediates. Initial cleavages occurred at distances between 10 and 44 amino acids from the N- or C-terminus of the molecule and often involved removal of a fragment from both the N- and C-termini of RCM-lysozyme. Our data indicate that degradation of proteins by MPCs from higher organisms involves a nonprocessive mechanism comprised of multiple, independent cleavages with dissociation of degradation intermediates. A general model for protein degradation by the MPC is discussed.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Wang R,Chait BT,Wolf I,Kohanski RA,Cardozo C

doi

10.1021/bi990826h

subject

Has Abstract

pub_date

1999-11-02 00:00:00

pages

14573-81

issue

eissn

0006-2960

issn

1520-4995

pii

bi990826h

journal_volume

pub_type

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Lysozyme degradation by the bovine multicatalytic proteinase complex (proteasome): evidence for a nonprocessive mode of degradation.