The three-dimensional structure at 2.4 Å resolution of glycosylated proteinase A from the lysosome-like vacuole of Saccharomyces cerevisiae

Aguilar, Carlos Fernando; Cronin, Nora; Badasso, M.; Dreyer, Thomas; Newman, Matthew; Cooper, J.B.; Hoover, Dennis J.; Wood, Stephen P.; Johnson, Mark S.; Blundell, Tom L.

doi:10.1006/jmbi.1996.0880

Cited by 35 publications

(21 citation statements)

References 68 publications

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“…A Bipartite Signal Targets Misfolded Glycoproteins to ERAD CPY* and PrA* are well-studied model Htm1p-dependent ERAD substrates with known crystal structures for their folded counterparts carboxypeptidase Y (CPY) and proteinase A (PrA; Finger et al, 1993;Endrizzi et al, 1994;Aguilar et al, 1997). The prc1-1 (CPY*) allele contains a missense mutation that results in the G255R change at the core of the folded protein.…”

Section: Resultsmentioning

confidence: 99%

Intrinsic Conformational Determinants Signal Protein Misfolding to the Hrd1/Htm1 Endoplasmic Reticulum–associated Degradation System

Xie

Kanehara

Sayeed

et al. 2009

MBoC

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Endoplasmic reticulum (ER) quality control mechanisms monitor the folding of nascent polypeptides of the secretory pathway. These are dynamic processes that retain folding proteins, promote the transport of conformationally mature proteins, and target misfolded proteins to ER-associated degradation (ERAD) pathways. Aided by the identification of numerous ERAD factors, late functions that include substrate extraction, ubiquitination, and degradation are fairly well described. By contrast, the mechanisms of substrate recognition remain mysterious. For some substrates, a specific N-linked glycan forms part of the recognition code but how it is read is incompletely understood. In this study, systematic analysis of model substrates revealed such glycans mark structural determinants that are sensitive to the overall folding state of the molecule. This strategy effectively generates intrinsic folding sensors that communicate with high fidelity to ERAD. Normally, these segments fold into the mature structure to pass the ERAD checkpoint. However, should a molecule fail to fold completely, they form a bipartite signal that comprises the unfolded local structure and adjacent enzymatically remodeled glycan. Only if both elements are present will the substrate be targeted to the ERAD pathway for degradation. INTRODUCTIONQuality control pathways monitor protein folding and assembly throughout the cell. Although generally poorly understood, the best-described systems are those of the endoplasmic reticulum (ER). ER quality control (ERQC) describes an assortment of mechanisms that, collectively, retains unfolded or unassembled proteins and targets irreversibly misfolded proteins for destruction through the appropriate ERassociated degradation (ERAD) pathway.Newly synthesized secretory and membrane proteins translocate across the ER membrane through the Sec61 translocon pore. The narrow pore size restricts passage to unfolded proteins so factors needed for their maturation reside in the ER (Rapoport, 2007). For most molecules, ER quality control serves to prevent their transport during the folding process. Once folding is complete, intrinsic export signals guide their transport via cargo sorting receptors concentrated at and in the membranes of ER exit sites marked by COPII coat proteins (Sato and Nakano, 2007).Proteins that fail to fold are targeted to ERAD for turnover. Metazoans maintain multiple ERAD pathways with the total number unknown (Nakatsukasa and Brodsky, 2008). The best understood is linked to the calnexin/calreticulin folding cycle (for review, see Helenius and Aebi, 2004). Newly synthesized glycoproteins bind calnexin or calreticulin via mono-glucosylated N-glycans. The interaction assists in the folding process by preventing aggregation and by engaging other factors. Proteins failing to fold are directed to ERAD via the action of the ER degradationenhancing ␣-mannosidase-like protein (EDEM; Molinari et al., 2003;Oda et al., 2003). Recently, EDEM1 and EDEM3 overexpression was shown to accelerate the demannosylatio...

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Section: Resultsmentioning

confidence: 99%

Intrinsic Conformational Determinants Signal Protein Misfolding to the Hrd1/Htm1 Endoplasmic Reticulum–associated Degradation System

Xie

Kanehara

Sayeed

et al. 2009

MBoC

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“…Cathepsin D is a lysosomal aspartic protease that is known to mediate cell death processes in tissue culture cells [12]. PEP4 encodes a vacuolar aspartic endopeptidase that is homologous to vertebrate cathepsin D [32]. Thus, we tested whether Pep4p was required for nucleoporin proteolysis in H 2 O 2 -treated cells.…”

Section: Nucleoporin Proteolysis Is Dependent On the Cathepsin D Homomentioning

confidence: 99%

Increased nuclear envelope permeability and Pep4p-dependent degradation of nucleoporins during hydrogen peroxide-induced cell death

Mason¹,

Shul'ga²,

Undavai³

et al. 2005

FEMS Yeast Research

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The death of yeast treated with hydrogen peroxide (H(2)O(2)) shares a number of morphological and biochemical features with mammalian apoptosis. In this study, we report that the permeability of yeast nuclear envelopes (NE) increased during H(2)O(2)-induced cell death. Similar phenomena have been observed during apoptosis in mammalian tissue culture cells. Increased NE permeability in yeast was temporally correlated with an increase in the production of reactive-oxygen species (ROS). Later, after ROS levels began to decline and viability was lost, specific nuclear pore complex (NPC) proteins (nucleoporins) were degraded. Although caspases are responsible for the degradation of mammalian nucleoporins during apoptosis, the deletion of the metacaspase gene YCA1 had no effect on the stability of yeast nucleoporins. Instead, Pep4p, a vacuolar cathepsin D homolog, was responsible for the proteolysis of nucleoporins. Coincident with nucleoporin degradation, a Pep4p-EGFP reporter migrated out of the vacuole in H(2)O(2)-treated cells. We conclude that increases in ROS and NPC permeability occur relatively early during H(2)O(2)-induced cell death. Later, Pep4p migrates out of vacuoles and degrades nucleoporins after the cells are effectively dead.

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“…Generally, the active enzyme consists of a single peptide chain of about 320-360 amino acid residues and has a molecular mass of 32-36 kDa. X-ray crystallographic analyses of various aspartic proteinases showed that they are composed of β-strand secondary structures arranged in a bilobal conformation (Aguilar et al 1997;Claverie-Martin and Vega-Hernandez 2007). The two lobes are homologous of each other and have most likely evolved by gene duplication.…”

mentioning

confidence: 99%

Aspartic proteinases from Mucor spp. in cheese manufacturing

Yeğin

Fernández‐Lahore

Salgado

et al. 2010

Appl Microbiol Biotechnol

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Filamentous fungi belonging to the order of Mucorales are well known as producers of aspartic proteinases depicting milk-clotting activity. The biosynthesis level, the biochemical characteristics, and the technological properties of the resulting proteinases are affected by the producer strain and the mode of cultivation. While the milk-clotting enzymes produced by the Rhizomucor spp. have been extensively studied in the past, much less is known on the properties and potential applications of the aspartic proteinases obtained for Mucor spp. Indeed, several Mucor spp. strains have been reported as a potential source of milk-clotting enzymes having unique technological properties. Both submerged fermentation and solid substrate cultivation are proven alternatives for the production of Mucor spp. aspartic proteinases. This review provides an overview on the bioprocessing routes to obtain large amounts of these enzymes, on their structural characteristics as related to their functional properties, and on their industrial applications with focus on cheese manufacturing.

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The three-dimensional structure at 2.4 Å resolution of glycosylated proteinase A from the lysosome-like vacuole of Saccharomyces cerevisiae

Cited by 35 publications

References 68 publications

Intrinsic Conformational Determinants Signal Protein Misfolding to the Hrd1/Htm1 Endoplasmic Reticulum–associated Degradation System

Intrinsic Conformational Determinants Signal Protein Misfolding to the Hrd1/Htm1 Endoplasmic Reticulum–associated Degradation System

Increased nuclear envelope permeability and Pep4p-dependent degradation of nucleoporins during hydrogen peroxide-induced cell death

Aspartic proteinases from Mucor spp. in cheese manufacturing

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