Identification of a Yeast Peroxisomal Member of the Family of AMP‐Binding Proteins

Blobel, Fabian; Erdmann, Ralf

doi:10.1111/j.1432-1033.1996.0468h.x

Cited by 67 publications

(60 citation statements)

References 50 publications

(52 reference statements)

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“…Equal proportions of pellet (P) and supernatant (S) fractions were analyzed by SDS-PAGE and Western blot analysis using specific antibodies against Idp3p, the integral membrane protein Pex3p (34), Pcs60p (32), and the peroxisomal matrix marker Fox3p (30). As Idp3p is hardly extracted by high salt treatment and totally extracted at pH 11, it behaves like Pcs60p, which is localized in the peroxisomal matrix but is also found loosely associated with the peroxisomal membrane (32). B, protease protection analysis of purified peroxisomes.…”

Section: Discussionmentioning

confidence: 99%

“…Idp3p also does not seem to be tightly associated with the peroxisomal membrane, since part of the protein could be extracted with high salt. The extraction properties of Idp3p are similar to those observed for Pcs60p, a protein of the peroxisomal matrix that is also loosely associated with the peroxisomal membrane (32). To distinguish whether Idp3p is associated with the outer aspects of peroxisomes or whether the protein resides in the peroxisomal lumen, we analyzed the sensitivity of organellar Idp3p to externally added proteases.…”

Section: Peroxisomes Of S Cerevisiae Contain An Nadp-dependentmentioning

confidence: 99%

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IDP3 Encodes a Peroxisomal NADP-dependent Isocitrate Dehydrogenase Required for the β-Oxidation of Unsaturated Fatty Acids

Henke

Girzalsky

Berteaux‐Lecellier

et al. 1998

Journal of Biological Chemistry

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106

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In Saccharomyces cerevisiae the metabolic degradation of saturated fatty acids is exclusively confined to peroxisomes. In addition to a functional ␤-oxidation system, the degradation of unsaturated fatty acids requires auxiliary enzymes, including a ⌬2,⌬3-enoyl-CoA isomerase and an NADPH-dependent 2,4-dienoyl-CoA reductase. We found both enzymes to be present in yeast peroxisomes. The impermeability of the peroxisomal membrane for pyrimidine nucleotides led to the question of how the NADPH needed by the reductase is regenerated in the peroxisomal lumen. We report the identification and functional analysis of the IDP3 gene product, which is a yeast peroxisomal NADP-dependent isocitrate dehydrogenase. The newly identified peroxisomal protein is homologous to the mitochondrial Idp1p and cytosolic Idp2p, which both are yeast NADPdependent isocitrate dehydrogenases. Yeast cells lacking Idp3p grow normally on saturated fatty acids, but growth is impaired on unsaturated fatty acids, indicating that the peroxisomal Idp3p is involved in their metabolic utilization. The data presented are consistent with the assumption that peroxisomes of S. cerevisiae contain the enzyme equipment needed for the degradation of unsaturated fatty acids, including an NADPdependent isocitrate dehydrogenase, a putative constituent of a peroxisomal NADPH-regenerating redox system.

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Section: Discussionmentioning

confidence: 99%

Section: Peroxisomes Of S Cerevisiae Contain An Nadp-dependentmentioning

confidence: 99%

IDP3 Encodes a Peroxisomal NADP-dependent Isocitrate Dehydrogenase Required for the β-Oxidation of Unsaturated Fatty Acids

Henke

Girzalsky

Berteaux‐Lecellier

et al. 1998

Journal of Biological Chemistry

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106

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“…Finally, in addition to the 10 major clades, an outlier containing a single yeast protein (sc6) was noted. This protein, known as Pcs60p or Fatp2, is a peroxisomal protein predicted to have ACS activity (46). We previously reported that Fatp2 belonged to an ACS subfamily that contains fungal, plant, and bacterial, but no mammalian, enzymes (11).…”

Section: Phylogenetic Analyses Of Human and Nonhuman Acs Sequencesmentioning

confidence: 99%

Evidence for 26 distinct acyl-coenzyme A synthetase genes in the human genome

Watkins

Maiguel

Jia

et al. 2007

Journal of Lipid Research

301

328

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Acyl-coenzyme A synthetases (ACSs) catalyze the fundamental, initial reaction in fatty acid metabolism. "Activation" of fatty acids by thioesterification to CoA allows their participation in both anabolic and catabolic pathways. The availability of the sequenced human genome has facilitated the investigation of the number of ACS genes present. Using two conserved amino acid sequence motifs to probe human DNA databases, 26 ACS family genes/proteins were identified. ACS activity in either humans or rodents was demonstrated previously for 20 proteins, but 6 remain candidate ACSs. For two candidates, cDNA was cloned, protein was expressed in COS-1 cells, and ACS activity was detected. Amino acid sequence similarities were used to assign enzymes into subfamilies, and subfamily assignments were consistent with acyl chain length preference. Four of the 26 proteins did not fit into a subfamily, and bootstrap analysis of phylograms was consistent with evolutionary divergence. Three additional conserved amino acid sequence motifs were identified that likely have functional or structural roles. The existence of many ACSs suggests that each plays a unique role, directing the acyl-CoA product to a specific metabolic fate. Knowing the full complement of ACS genes in the human genome will facilitate future studies to characterize their specific biological functions.-Watkins, P. A., D. Maiguel, Z. Jia, and J. Pevsner. Evidence for 26 distinct acyl-coenzyme A synthetase genes in the human genome. J. Lipid Res.

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“…The antibodies used were obtained from commercial sources or described previously, namely rabbit anti-Pex13p (Girzalsky et al, 1999), anti-Pex11p (Erdmann and Blobel, 1995), anti-Pcs60p (Blobel and Erdmann, 1996), and anti-green fluorescent protein (GFP; W.H. Kunau, Bochum, Germany) as well Full-length PEX13 and various truncations thereof were fused to the GAL4 binding domain (Gal4p-BD) in vector pPC97.…”

Section: Antibodies and Western Blottingmentioning

confidence: 99%

Peroxisomal Membrane Proteins Contain Common Pex19p-binding Sites that Are an Integral Part of Their Targeting Signals

Rottensteiner

Kramer

Lorenzen

et al. 2004

MBoC

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161

196

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Targeting of peroxisomal membrane proteins (PMPs) is a multistep process that requires not only recognition of PMPs in the cytosol but also their insertion into the peroxisomal membrane. As a consequence, targeting signals of PMPs (mPTS) are rather complex. A candidate protein for the PMP recognition event is Pex19p, which interacts with most PMPs. However, the respective Pex19p-binding sites are ill-defined and it is currently disputed whether these sites are contained within mPTS. By using synthetic peptide scans and yeast two-hybrid analyses, we determined and characterized Pex19p-binding sites in Pex11p and Pex13p, two PMPs from Saccharomyces cerevisiae. The sites turned out to be composed of a short helical motif with a minimal length of 11 amino acids. With the acquired data, it proved possible to predict and experimentally verify Pex19p-binding sites in several other PMPs by applying a pattern search and a prediction matrix. A peroxisomally targeted Pex13p fragment became mislocalized to the endoplasmic reticulum in the absence of its Pex19p-binding site. By adding the heterologous binding site of Pex11p, peroxisomal targeting of the Pex13p fragment was restored. We conclude that Pex19p-binding sites are well-defined entities that represent an essential part of the mPTS. INTRODUCTIONPeroxisomes are ubiquitous organelles of eukaryotic cells, whose proteins are imported posttranslationally. Matrix proteins are directed to peroxisomes by either of two targeting signals, a C-terminal PTS1 or an N-terminal PTS2. The topogenesis of peroxisomal membrane proteins (PMPs) is accomplished by yet another mechanism, because most of the peroxin mutants, characterized by their defect in the biogenesis of peroxisomes, exhibit a block in matrix protein import, but do import PMPs normally (Lazarow and Fujiki, 1985;Gould and Valle, 2000;Subramani et al., 2000;Purdue and Lazarow, 2001;Eckert and Erdmann, 2003). To date, only three peroxins with a potential role in PMP targeting have been identified, namely Pex3p (Hettema et al., 2000;South et al., 2000), Pex16p in mammals (South and Gould, 1999;Honsho et al., 2002), and Pex19p (Gö tte et al., 1998;Matsuzono et al., 1999;Snyder et al., 1999;Soukupova et al., 1999). In cells lacking any of these proteins, PMPs are either degraded or mistargeted to other subcellular compartments such as mitochondria, the endoplasmic reticulum (ER) and membranes of unknown origin (Ghaedi et al., 2000;Hettema et al., 2000;Sacksteder et al., 2000).In accordance with a distinct pathway, PMPs use neither PTS1 nor PTS2. The targeting signals of PMPs (mPTS) that direct and insert PMPs into the peroxisomal membrane have been determined for a number of PMPs of several species. Despite some differences, a picture emerged from these studies of a targeting signal consisting of one or more transmembrane domains in conjunction with a short sequence, which contains either a cluster of basic residues or a mixture of basic and hydrophobic amino acids (Dyer et al., 1996;Baerends et al., 2000b;Pause et al., 2000;...

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Identification of a Yeast Peroxisomal Member of the Family of AMP‐Binding Proteins

Cited by 67 publications

References 50 publications

IDP3 Encodes a Peroxisomal NADP-dependent Isocitrate Dehydrogenase Required for the β-Oxidation of Unsaturated Fatty Acids

IDP3 Encodes a Peroxisomal NADP-dependent Isocitrate Dehydrogenase Required for the β-Oxidation of Unsaturated Fatty Acids

Evidence for 26 distinct acyl-coenzyme A synthetase genes in the human genome

Peroxisomal Membrane Proteins Contain Common Pex19p-binding Sites that Are an Integral Part of Their Targeting Signals

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