Proteasome from Thermoplasma acidophilum: a threonine protease.

The catalytic mechanism of the 20S proteasome from the archaebacterium Thermoplasma acidophilum has been analyzed by site-directed mutagenesis of the beta subunit and by inhibitor studies. Deletion of the amino-terminal threonine or its mutation to alanine led to inactivation of the enzyme. Mutation of the residue to serine led to a fully active enzyme, which was over ten times more sensitive to the serine protease inhibitor 3,4-dichloroisocoumarin. In combination with the crystal structure of a proteasome-inhibitor complex, the data show that the nucleophilic attack is mediated by the amino-terminal threonine of processed beta subunits. The conservation pattern of this residue in eukaryotic sequences suggests that at least three of the seven eukaryotic beta-type subunit branches should be proteolytically inactive.

[1]  R. Mason.  Characterization of the active site of human multicatalytic proteinase. , 1990, The Biochemical journal.

[2]  W. Baumeister,et al.  Structural features of the 26 S proteasome complex. , 1993, Journal of molecular biology.

[3]  A. Goldberg,et al.  Proteolysis, proteasomes and antigen presentation , 1992, Nature.

[4]  P. Kloetzel,et al.  20 S proteasomes are assembled via distinct precursor complexes. Processing of LMP2 and LMP7 proproteins takes place in 13-16 S preproteasome complexes. , 1994, Journal of molecular biology.

[5]  C. Slaughter,et al.  The primary structures of four subunits of the human, high-molecular-weight proteinase, macropain (proteasome), are distinct but homologous. , 1991, Biochimica et biophysica acta.

[6]  M Orlowski,et al.  Evidence for the presence of five distinct proteolytic components in the pituitary multicatalytic proteinase complex. Properties of two components cleaving bonds on the carboxyl side of branched chain and small neutral amino acids. , 1993, Biochemistry.

[7]  A. Goldberg,et al.  Inhibitors of the proteasome block the degradation of most cell proteins and the generation of peptides presented on MHC class I molecules , 1994, Cell.

[8]  R Hegerl,et al.  The three‐dimensional structure of proteasomes from Thermoplasma acidophilum as determined by electron microscopy using random conical tilting , 1991, FEBS letters.

[9]  F. Studier,et al.  Use of T7 RNA polymerase to direct expression of cloned genes. , 1990, Methods in enzymology.

[10]  J. Kleinz,et al.  Critical elements in proteasome assembly , 1994, Nature Structural Biology.

[11]  J. Kleinschmidt,et al.  Proteinase yscE, the yeast proteasome/multicatalytic‐multifunctional proteinase: mutants unravel its function in stress induced proteolysis and uncover its necessity for cell survival. , 1991, The EMBO journal.

[12]  W. Baumeister,et al.  Localization of a sequence motif complementary to the nuclear localization signal in proteasomes from Thermoplasma acidophilum by immunoelectron microscopy. , 1992, Journal of structural biology.

[13]  A. Cornish-Bowden,et al.  The direct linear plot. A new graphical procedure for estimating enzyme kinetic parameters. , 1974, The Biochemical journal.

[14]  Eleanor J. Dodson,et al.  Penicillin acylase has a single-amino-acid catalytic centre , 1996, Nature.

[15]  A. Rivett,et al.  Use of serine-protease inhibitors as probes for the different proteolytic activities of the rat liver multicatalytic proteinase complex. , 1992, European journal of biochemistry.

[16]  P. A. Peterson,et al.  Alternative exon usage and processing of the major histocompatibility complex-encoded proteasome subunits. , 1992, The Journal of biological chemistry.

[17]  C. Enenkel,et al.  PRE3, highly homologous to the human major histocompatibility complex‐linked LMP2 (RING12) gene, codes for a yeast proteasome subunit necessary for the peptidylglutamyl‐peptide hydrolyzing activity , 1994, FEBS letters.

[18]  R Hegerl,et al.  Subunit stoichiometry and three‐dimensional arrangement in proteasomes from Thermoplasma acidophilum. , 1992, The EMBO journal.

[19]  M Karplus,et al.  Analysis of side-chain orientations in homologous proteins. , 1987, Journal of molecular biology.

[20]  M. Rechsteiner.  Natural substrates of the Ubiquitin proteolytic pathway , 1991, Cell.

[21]  K. Tanaka,et al.  Molecular cloning and sequence analysis of cDNAs for five major subunits of human proteasomes (multi-catalytic proteinase complexes). , 1991, Biochimica et biophysica acta.

[22]  M. Orłowski,et al.  A 3,4-dichloroisocoumarin-resistant component of the multicatalytic proteinase complex. , 1992, Biochemistry.

[23]  R. Schoepfer.  The pRSET family of T7 promoter expression vectors for Escherichia coli. , 1993, Gene.

[24]  A. Gruhler,et al.  The PRE4 gene codes for a subunit of the yeast proteasome necessary for peptidylglutamyl-peptide-hydrolyzing activity. Mutations link the proteasome to stress- and ubiquitin-dependent proteolysis. , 1993, The Journal of biological chemistry.

[25]  W. Baumeister,et al.  The proteasome from Thermoplasma acidophilum is neither a cysteine nor a serine protease , 1995, FEBS letters.

[26]  S. Wilk,et al.  Cation‐Sensitive Neutral Endopeptidase: Isolation and Specificity of the Bovine Pituitary Enzyme , 1980, Journal of neurochemistry.

[27]  A. Goldberg,et al.  Gamma-interferon and expression of MHC genes regulate peptide hydrolysis by proteasomes. , 1993, Nature.

[28]  W. Baumeister,et al.  Thermoplasma acidophilum proteasomes degrade partially unfolded and ubiquitin‐associated proteins , 1993, FEBS letters.

[29]  P M Kloetzel,et al.  Interferon gamma stimulation modulates the proteolytic activity and cleavage site preference of 20S mouse proteasomes , 1994, The Journal of experimental medicine.

[30]  W. Baumeister,et al.  Conformational constraints in protein degradation by the 20S proteasome , 1995, Nature Structural Biology.

[31]  E. Shaw,et al.  Reaction of proteasomes with peptidylchloromethanes and peptidyldiazomethanes. , 1993, The Biochemical journal.

[32]  W. Baumeister,et al.  Molecular characterization of the "26S" proteasome complex from rat liver. , 1993, Journal of structural biology.

[33]  A. Ciechanover,et al.  The ubiquitin system for protein degradation. , 1992, Annual review of biochemistry.

[34]  W. Baumeister,et al.  Proteasomes from Dictyostelium discoideum: characterization of structure and function. , 1993, Journal of structural biology.

[35]  W. Baumeister,et al.  Proteasome sequences in eubacteria. , 1994, Trends in biochemical sciences.

[36]  W. Baumeister,et al.  Biochemical properties of the proteasome from Thermoplasma acidophilum. , 1992, European journal of biochemistry.

[37]  W. Baumeister,et al.  The multicatalytic proteinase (prosome) is ubiquitous from eukaryotes to archaebacteria , 1989, FEBS letters.

[38]  W. Baumeister,et al.  Expression of functional Thermoplasma acidophilum proteasomes in Escherichia coli , 1992, FEBS letters.

[39]  D. Finley,et al.  MHC-linked LMP gene products specifically alter peptidase activities of the proteasome , 1993, Nature.

[40]  A. Goldberg.  The mechanism and functions of ATP-dependent proteases in bacterial and animal cells. , 1992, European journal of biochemistry.

[41]  J. Monaco,et al.  MHC-linked low-molecular mass polypeptide subunits define distinct subsets of proteasomes. Implications for divergent function among distinct proteasome subsets. , 1993, Journal of immunology.

[42]  W. Baumeister,et al.  Electron microscopy and image analysis of the multicatalytic proteinase , 1988, FEBS letters.

[43]  P. A. Peterson,et al.  Proteasomes are regulated by interferon gamma: implications for antigen processing. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[44]  K. Tanaka,et al.  Interferon-gamma induces different subunit organizations and functional diversity of proteasomes. , 1994, Journal of biochemistry.

[45]  K. Kobashi,et al.  The quaternary structure of carp muscle aklaline protease. , 1980, Biochimica et biophysica acta.

[46]  M. Orłowski,et al.  A multicatalytic protease complex from pituitary that forms enkephalin and enkephalin containing peptides. , 1981, Biochemical and biophysical research communications.