Recognition of a Functional Peroxisome Type 1 Target by the Dynamic Import Receptor Pex5p

Stanley, Will A.; Filipp, Fabian V.; Kursula, Petri; Schüller, Nicole; Erdmann, Ralf; Schliebs, Wolfgang; Sattler, Michael; Wilmanns, Matthias

doi:10.1016/j.molcel.2006.10.024

Cited by 155 publications

(243 citation statements)

References 60 publications

(64 reference statements)

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“…Each of the four variants, regardless of the presence or absence of either presequence or PTS1, were found by far-UV CD to be natively folded, as estimated by far-UV CD and confirming previous structural data [3,13,26,[29][30][31]. Previous spectroscopic studies [32,33] have shown that the presequence of SCP2 does not display regular secondary structure-a conclusion confirmed by the CD data presented here.…”

Section: Discussionsupporting

confidence: 90%

“…Previous spectroscopic studies [32,33] have shown that the presequence of SCP2 does not display regular secondary structure-a conclusion confirmed by the CD data presented here. In contrast to one earlier CD study [34], in which the presence of the presequence was found to significantly reduce the ordered secondary structure content of SCP2, our data are consistent with available NMR data [18,31,32] indicating that the overall fold of preSCP2 and mSCP2, aside from the coiled presequence, does not change. We additionally demonstrate that removal of the PTS1 tripeptide does not alter SCP2 secondary structure and thus is not critical for SCP2 folding, as could be expected from high-resolution structural analyses Fig.…”

Section: Discussionsupporting

confidence: 88%

“…Note the increase in relative abundance of apo-preSCP2 (P peaks) with increasing CV. [26,30,31] in which the C-terminal segment of SCP2 is found to be in a coiled conformation. Crystal structures of liganded SCP2 [13,29] show the C-terminal segment adopting an a-helical structure ''capping'' the ligand binding pocket.…”

Section: Discussionmentioning

confidence: 98%

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Investigation of the ligand spectrum of human sterol carrier protein 2 using a direct mass spectrometry assay

Stanley

Versluis

Schultz

et al. 2007

Archives of Biochemistry and Biophysics

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Sterol carrier protein 2 (SCP2) has been investigated by nearly native electrospray ionisation mass spectrometry in the presence of long chain fatty acyl CoAs (LCFA-CoAs) and carnitine derivatives of equivalent fatty acid chain length (LCFA-carnitines). Four SCP2 constructs were compared to examine the influence of the N-terminal presequence and the C-terminal peroxisomal targeting signal on ligand binding. Removal of N-or C-terminal residues did not influence ligand binding. The observation that LCFA-CoAs are high affinity ligands for SCP2 was confirmed, while LCFA-carnitines were demonstrated for the first time not to interact with SCP2. LCFACoAs formed non-covalent complexes with SCP2 of 2:1 and 1:1 stoichiometry, which could be dissociated by elevating the energy of the ions upon entrance to the mass spectrometer. A fluorescence-competition assay using Nile Red butyric acid confirmed the mass spectrometric observations in solution. The physiological significance of the lack of LCFA-carnitine binding by SCP2 is discussed.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionsupporting

confidence: 88%

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Investigation of the ligand spectrum of human sterol carrier protein 2 using a direct mass spectrometry assay

Stanley

Versluis

Schultz

et al. 2007

Archives of Biochemistry and Biophysics

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“…Relaxation times were measured in an interleaved fashion with 12-14 relaxation delays for T 1 15 N T 1ρ relaxation data for SCP2 and SCP2-Pex5p were measured as described (26).…”

Section: Relaxation Analysismentioning

confidence: 99%

Conformational Plasticity of the Lipid Transfer Protein SCP2

Filipp

Sattler

2007

Biochemistry

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The nonspecific lipid transfer protein sterol carrier protein 2 (SCP2) is involved in organellar fatty acid metabolism. A hydrophobic cavity in the structure of SCP2 accommodates a wide variety of apolar ligands such as cholesterol derivatives or fatty acyl-coenzyme A (CoA) conjugates. The properties of this nonspecific lipid binding pocket are explored using NMR chemical shift perturbations, paramagnetic relaxation enhancement, amide hydrogen exchange, and 15 N relaxation measurements. A common binding cavity shared by different physiological ligands is identified. NMR relaxation measurements reveal that residues in the three C-terminal α-helices within the lipid binding region exhibit mobility at fast (picosecond to nanosecond) and slow (microsecond to millisecond) time scales. Ligand binding is associated with a considerable loss of peptide backbone mobility. The observed conformational dynamics in SCP2 may play a role for the access of hydrophobic ligands to an occluded binding pocket. The C-terminal peroxisomal targeting signal of SCP2 is specifically recognized by the Pex5p receptor protein, which conducts cargo proteins toward the peroxisomal organelle. Neither the C-terminal targeting signal nor the N-terminal precursor sequence interferes with lipid binding by SCP2. The α-helices involved in lipid binding also mediate a secondary interaction interface with the Pex5p receptor. Silencing of conformational dynamics of the peptide backbone in these helices upon either lipid or Pex5p binding might communicate the loading state of the cargo protein to the targeting receptor. Graphical Abstract HHS Public AccessAuthor manuscript Biochemistry. Author manuscript; available in PMC 2016 September 08. Author Manuscript Author ManuscriptAuthor Manuscript Author ManuscriptLipid transfer proteins are auxiliary shuttles ensuring the appropriate localization of lipids in cellular subcompartments. Sterol carrier protein 2 (SCP2 1 ) has been reported to interact with a wide variety of lipids associated with peroxisomal metabolism, including acyl-CoA conjugates as high-affinity ligands (1-3). Disruption of the scp2 gene in mice suggests bile acids (soluble derivatives of cholesterol) and branched-chain fatty acid CoAs as physiological ligands to be functionally associated with SCP2 (4-6). Enzymes involved in both cholesterol secretion and fatty acid oxidation include the peroxisomal thiolase, acylCoA hydrogenase, and enoyl-CoA hydratase (7,8), all of which are expressed as fusion proteins with a sterol carrier protein domain fold (9). The transfer of fatty acyl-CoA substrates by SCP2 may have evolved to assist enzymes involved in branched chain fatty acid α-oxidation or very long chain fatty acid β-oxidation in the peroxisome (8). In this context SCP2 may play a role in substrate and/or product binding during peroxisomal bile acid synthesis and fatty acid oxidation.Some molecular insight into the ligand binding site of the sterol carrier protein fold has been obtained from structural high-resolution studies of SC...

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“…The PTS1 sequence is recognized by the receptor protein Pex5p 8, 9. Pex5p recruits cargo proteins to the peroxisomal matrix by means of a C‐terminal tetratricopeptide repeat domain, which interacts with the PTS1 sequence 10. The N‐terminal region of Pex5p is required for membrane docking and recycling.…”

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confidence: 99%

Hansenula polymorpha Aat2p is targeted to peroxisomes via a novel Pex20p‐dependent pathway

et al. 2018

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Saccharomyces cerevisiae Aat2p contains a peroxisomal targeting signal type‐1 and localizes to peroxisomes in oleate‐grown cells, but not in glucose‐grown cells. Here, we have investigated Aat2p from the yeast Hansenula polymorpha, which lacks a recognizable peroxisomal targeting signal. Aat2p tagged with GFP at its C terminus displays a dual cytosol‐peroxisome localization in ethanol‐grown cells. The partial peroxisomal localization of Aat2p persisted in the absence of the classical cycling receptors Pex5p and Pex7p but Aat2p targeting to peroxisomes was reduced in cells deleted for the matrix protein import factors PEX1, PEX2 and PEX13. Furthermore, we demonstrate that Aat2p targeting to peroxisomes requires Pex20p. Together, our data identify a Pex20p‐dependent pathway for targeting Aat2p to peroxisomes.

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Recognition of a Functional Peroxisome Type 1 Target by the Dynamic Import Receptor Pex5p

Cited by 155 publications

References 60 publications

Investigation of the ligand spectrum of human sterol carrier protein 2 using a direct mass spectrometry assay

Investigation of the ligand spectrum of human sterol carrier protein 2 using a direct mass spectrometry assay

Conformational Plasticity of the Lipid Transfer Protein SCP2

Hansenula polymorpha Aat2p is targeted to peroxisomes via a novel Pex20p‐dependent pathway

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