A selective tagging method for detecting minor alternatively-localized populations of a protein is used to study a disease-associated transmembrane form of prion protein. The analysis reveals key features of transmembrane prion protein metabolism and one way this is altered by human disease-causing mutants

Addya S.

Aguzzi A.

Amy B. Emerman

Anandatheerthavarada H. K.

Ashok A.

Beckett D.

Boopathi E.

Chakrabarti O.

Colombo S.

Dasari V. R.

Drisaldi B.

Fons R. D.

Goder V.

Goulet B.

Hegde R. S.

Howarth M.

Jungnickel B.

Kang S. W.

Kim S. J.

Krieg U. C.

Kulman J. D.

Kurzchalia T. V.

Land T.

Lorenz H.

Lu N. Z.

Ma J.

Matthews G. D.

Mize G. J.

Naamati A.

Negro A.

Oishee Chakrabarti

Pop C.

Prusiner S. B.

Ramanujan S. Hegde

Rane N. S.

Regev-Rudzki N.

Reid Gilmore

Robin M. A.

Sannerud R.

Sass E.

Schatz P. J.

Shaffer K. L.

Sharma A.

Stein I.

Stewart R. S.

Tong W. H.

Westergard L.

Yang W.

Yogev O.

Yudt M. R.

Zai-Rong Zhang

English

PubMed

 Proteins are often made in more than one form, with alternate versions sometimes residing in different cellular compartments than the primary species. The mammalian prion protein (PrP), a cell surface GPI-anchored protein, is a particularly noteworthy example for which minor cytosolic and transmembrane forms have been implicated in disease pathogenesis. To study these minor species, we used a selective labeling strategy in which spatially restricted expression of a biotinylating enzyme was combined with asymmetric engineering of the cognate acceptor sequence into PrP. Using this method, we could show that even wild-type PrP generates small amounts of the CtmPrP transmembrane form. Selective detection of CtmPrP allowed us to reveal its N-terminal processing, long half-life, residence in both intracellular and cell surface locations, and eventual degradation in the lysosome. Surprisingly, some human disease-causing mutants in PrP selectively stabilized CtmPrP, revealing a previously unanticipated mechanism of CtmPrP up-regulation that may contribute to disease. Thus, spatiotemporal tagging has uncovered novel aspects of normal and mutant PrP metabolism and should be readily applicable to the analysis of minor topologic isoforms of other proteins. </jats:p

Crossref

Molecular Biology of the Cell

Compartment-Restricted Biotinylation Reveals Novel Features of Prion Protein Metabolism in Vivo

A cathepsin L isoform that is devoid of a signal peptide localizes to the nucleus in S phase and processes the CDP/Cux transcription factor.

A minimal peptide substrate in biotin holoenzyme synthetase-catalyzed biotinylation.

A new transgenic mouse model of Gerstmann-Straussler-Scheinker syndrome caused by the A117V mutation of PRNP.

A posttargeting signal sequence recognition event in the endoplasmic reticulum membrane.

A transmembrane form of the prion protein contains an uncleaved signal peptide and is retained in the endoplasmic Reticulum.

A transmembrane form of the prion protein in neurodegenerative disease.

A transmembrane form of the prion protein is localized in the Golgi apparatus of neurons.

A versatile system for site-speciﬁc enzymatic biotinylation and regulated expression of proteins in cultured mammalian cells. Protein Expr.

Bimodal protein targeting through activation of cryptic mitochondrial targeting signals by an inducible cytosolic endoprotease.

Bimodal targeting of microsomal CYP2E1 to mitochondria through activation of an N-terminal chimeric signal by cAMP-mediated phosphorylation.

Cellular phenotyping of secretory and nuclear prion proteins associated with inherited prion diseases.

Combinatorial control of prion protein biogenesis by the signal sequence and transmembrane domain.

Conversion of PrP to a self-perpetuating PrPSc-like conformation in the cytosol.

Cotranslational partitioning of nascent prion protein into multiple populations at the translocation channel.

Dual targeting of Nfs1 and discovery of its novel processing enzyme,

Eclipsed distribution: a phenomenon of dual targeting of protein and its signiﬁcance.

Fumarase: a mitochondrial metabolic enzyme and a cytosolic/nuclear component of the DNA damage response.

Functional depletion of mahogunin by cytosolically exposed prion protein contributes to neurodegeneration.

Human caspases: activation, speciﬁcity, and regulation.

In vitro dissection of protein translocation into the mammalian endoplasmic reticulum. Methods Mol.

In vivo kinetics of protein targeting to the endoplasmic reticulum determined by site-speciﬁc phosphorylation.

Mitochondrial and cytosolic isoforms of yeast fumarase are derivatives of a single translation product and have identical amino termini.

Mitochondrial targeting and a novel transmembrane arrest of Alzheimer’s amyloid precursor protein impairs mitochondrial function in neuronal cells.

Most pathogenic mutations do not alter the membrane topology of the prion protein.

Mutant PrP is delayed in its exit from the endoplasmic reticulum, but neither wild-type nor mutant PrP undergoes retrotranslocation prior to proteasomal degradation.

Mutational analysis of topological determinants in prion protein (PrP) and measurement of transmembrane and cytosolic PrP during prion infection.

N-myristoylation determines dual targeting of mammalian NADH-cytochrome b5 reductase to ER and mitochondrial outer membranes by a mechanism of kinetic partitioning.

Neurodegenerative illness in transgenic mice expressing a transmembrane form of the prion protein.

Neurotoxicity and neurodegeneration when PrP accumulates in the cytosol.

Photocrosslinking of the signal sequence of nascent preprolactin to the 54-kilodalton polypeptide of the signal recognition particle.

Prion protein biosynthesis and its emerging role in neurodegeneration.

Prions: protein aggregation and infectious diseases.

Protection from cytosolic prion protein toxicity by modulation of protein translocation.

Reduced translocation of nascent prion protein during ER stress contributes to neurodegeneration.

Regulated expression of active biotinylated G-protein coupled receptors in mammalian cells.

Regulation of protein compartmentalization expands the diversity of protein function.

Retrotranslocation of prion proteins from the endoplasmic reticulum by preventing GPI signal transamidation.

Role of protein kinase C-mediated protein phosphorylation in mitochondrial translocation of mouse CYP1A1, which contains a non-canonical targeting signal.

Selective processing and metabolism of disease-causing mutant prion proteins. PLoS Pathog.

Selective regulation of bone cell apoptosis by translational isoforms of the glucocorticoid receptor.

Signal sequence insufﬁciency contributes to neurodegeneration caused by transmembrane prion protein.

Signal sequences control gating of the protein translocation channel in a substrate-speciﬁc manner.

Subcellular compartmentalization of human Nfu, an iron-sulfur cluster scaffold protein, and its ability to assemble a [4Fe-4S] cluster.

Substrate-speciﬁc function of the translocon-associated protein complex during translocation across the ER membrane.

Substrate-speciﬁc translocational attenuation during ER stress deﬁnes a pre-emptive quality control pathway.

Targeting of a human iron-sulfur cluster assembly enzyme, nifs, to different subcellular compartments is regulated through alternative AUG utilization.

Targeting of NH2-terminal-processed microsomal protein to mitochondria: a novel pathway for the biogenesis of hepatic mitochondrial P450MT2.

Targeting quantum dots to surface proteins in living cells with biotin ligase.

The cellular prion protein (PrP(C)): its physiological function and role in disease.

The efﬁciency of protein compartmentalization into the secretory pathway.

The glucocorticoid receptor: coding a diversity of proteins and responses through a single gene.

The metabolism and imaging in live cells of the bovine prion protein in its native form or carrying single amino acid substitutions.

The prion’s elusive reason for being.

The signal sequence of nascent preprolactin interacts with the 54K polypeptide of the signal recognition particle.

The single translation product of the FUM1 gene (fumarase) is processed in mitochondria before being distributed between the cytosol and mitochondria in Saccharomyces cerevisiae.

Trafﬁcking, a key player in regulated intramembrane proteolysis.

Translational regulatory mechanisms generate N-terminal glucocorticoid receptor isoforms with unique transcriptional target genes.

Transmissible and genetic prion diseases share a common pathway of neurodegeneration.

Use of peptide libraries to map the substrate speciﬁcity of a peptide-modifying enzyme: a 13 residue consensus peptide speciﬁes biotinylation in Escherichia coli.

Weak mitochondrial targeting sequence determines tissue-speciﬁc subcellular localization of glutamine synthetase in liver and brain cells.

Yeast aconitase in two locations and two metabolic pathways: seeing small amounts is believing.

http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3002386

Compartment-Restricted Biotinylation Reveals Novel Features of Prion Protein Metabolism in Vivo

Abstract

Similar works

Full text

Available Versions

Crossref