scholarly journals The Vacuolar Targeting Signal of the 2S Albumin from Brazil Nut Resides at the C Terminus and Involves the C-Terminal Propeptide as an Essential Element

1996 ◽  
Vol 112 (3) ◽  
pp. 975-985 ◽  
Author(s):  
G. Saalbach ◽  
M. Rosso ◽  
U. Schumann
Keyword(s):  
Essential Element ◽  
Brazil Nut ◽  
2S Albumin ◽  
C Terminus ◽  
Targeting Signal ◽  
Vacuolar Targeting

Peroxisome targeting signal of rat liver acyl-coenzyme A oxidase resides at the carboxy terminus

1989 ◽  
Vol 9 (1) ◽  
pp. 83-91
Author(s):  
S Miyazawa ◽  
T Osumi ◽  
T Hashimoto ◽  
K Ohno ◽  
S Miura ◽  
...  
Keyword(s):  
Amino Acid ◽  
Rat Liver ◽  
Coenzyme A ◽  
Terminal Region ◽  
Proteinase K ◽  
Amino Acid Residues ◽  
C Terminus ◽  
Targeting Signal ◽  
Acyl Coenzyme A

To identify the topogenic signal of peroxisomal acyl-coenzyme A oxidase (AOX) of rat liver, we carried out in vitro import experiments with mutant polypeptides of the enzyme. Full-length AOX and polypeptides that were truncated at the N-terminal region were efficiently imported into peroxisomes, as determined by resistance to externally added proteinase K. Polypeptides carrying internal deletions in the C-terminal region exhibited much lower import activities. Polypeptides that were truncated or mutated at the extreme C terminus were totally import negative. When the five amino acid residues at the extreme C terminus were attached to some of the import-negative polypeptides, the import activities were rescued. Moreover, the C-terminal 199 and 70 amino acid residues of AOX directed fusion proteins with two bacterial enzymes to peroxisomes. These results are interpreted to mean that the peroxisome targeting signal of AOX residues at the C terminus and the five or fewer residues at the extreme terminus have an obligatory function in targeting. The C-terminal internal region also has an important role for efficient import, possibly through a conformational effect.

scholarly journals Determination of the functional elements within the vacuolar targeting signal of barley lectin.

1993 ◽  
Vol 5 (5) ◽  
pp. 587-596 ◽  
Author(s):  
J E Dombrowski ◽  
M R Schroeder ◽  
S Y Bednarek ◽  
N V Raikhel
Keyword(s):  
Functional Elements ◽  
Targeting Signal ◽  
Vacuolar Targeting

scholarly journals Sequence-specific, Golgi-dependent vacuolar targeting of castor bean 2S albumin

2003 ◽  
Vol 36 (5) ◽  
pp. 711-719 ◽  
Author(s):  
Joanna C. Brown ◽  
Nicholas A. Jolliffe ◽  
Lorenzo Frigerio ◽  
Lynne M. Roberts
Keyword(s):  
Castor Bean ◽  
2S Albumin ◽  
Vacuolar Targeting

scholarly journals C-terminal tripeptide Ser-Asn-Leu (SNL) of human D-aspartate oxidase is a functional peroxisome-targeting signal

1998 ◽  
Vol 336 (2) ◽  
pp. 367-371 ◽  
Author(s):  
Leen AMERY ◽  
Chantal BREES ◽  
Myriam BAES ◽  
Chiaki SETOYAMA ◽  
Retsu MIURA ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Amino Acid ◽  
Fluorescent Protein ◽  
Consensus Sequence ◽  
C Terminus ◽  
Targeting Signal ◽  
Fluorescent Pattern ◽  
Green Fluorescent ◽  
Positively Charged

The functionality of the C-terminus (Ser-Asn-Leu; SNL) of human d-aspartate oxidase, an enzyme proposed to have a role in the inactivation of synaptically released d-aspartate, as a peroxisome-targeting signal (PTS1) was investigated in vivoand in vitro. Bacterially expressed human d-aspartate oxidase was shown to interact with the human PTS1-binding protein, peroxin protein 5 (PEX5p). Binding was gradually abolished by carboxypeptidase treatment of the oxidase and competitively inhibited by a Ser-Lys-Leu (SKL)-containing peptide. After transfection of mouse fibroblasts with a plasmid encoding green fluorescent protein (GFP) extended by PKSNL (the C-terminal pentapeptide of the oxidase), a punctate fluorescent pattern was evident. The modified GFP co-localized with peroxisomal thiolase as shown by indirect immunofluorescence. On transfection in fibroblasts lacking PEX5p receptor, GFP–PKSNL staining was cytosolic. Peroxisomal import of GFP extended by PGSNL (replacement of the positively charged fourth-last amino acid by glycine) seemed to be slower than that of GFP–PKSNL, whereas extension by PKSNG abolished the import of the modified GFP. Taken together, these results indicate that SNL, a tripeptide not fitting the PTS1 consensus currently defined in mammalian systems, acts as a functional PTS1 in mammalian systems, and that the consensus sequence, based on this work and that of other groups, has to be broadened to (S/A/C/K/N)-(K/R/H/Q/N/S)-L.

scholarly journals Di-Leucine Signals Mediate Targeting of Tyrosinase and Synaptotagmin to Synaptic-like Microvesicles within PC12 Cells

1999 ◽  
Vol 10 (11) ◽  
pp. 3979-3990 ◽  
Author(s):  
Anastasiya D. Blagoveshchenskaya ◽  
Eric W. Hewitt ◽  
Daniel F. Cutler
Keyword(s):  
Plasma Membrane ◽  
Pc12 Cells ◽  
Brefeldin A ◽  
Adaptor Protein ◽  
Dependent Manner ◽  
Protein Traffic ◽  
C Terminus ◽  
Targeting Signal

One pathway in forming synaptic-like microvesicles (SLMV) involves direct budding from the plasma membrane, requires adaptor protein 2 (AP2) and is brefeldin A (BFA) resistant. A second route leads from the plasma membrane to an endosomal intermediate from which SLMV bud in a BFA-sensitive, AP3-dependent manner. Because AP3 has been shown to bind to a di-leucine targeting signal in vitro, we have investigated whether this major class of targeting signals is capable of directing protein traffic to SLMV in vivo. We have found that a di-leucine signal within the cytoplasmic tail of human tyrosinase is responsible for the majority of the targeting of HRP-tyrosinase chimeras to SLMV in PC12 cells. Furthermore, we have discovered that a Met-Leu di-hydrophobic motif within the extreme C terminus of synaptotagmin I supports 20% of the SLMV targeting of a CD4-synaptotagmin chimera. All of the traffic to the SLMV mediated by either di-Leu or Met-Leu is BFA sensitive, strongly suggesting a role for AP3 and possibly for an endosomal intermediate in this process. The differential reduction in SLMV targeting for HRP-tyrosinase and CD4-synaptotagmin chimeras by di-alanine substitutions or BFA treatment implies that different proteins use the two routes to the SLMV to differing extents.

Stability of the major allergen Brazil nut 2S albumin (Ber e 1) to physiologically relevant in vitro gastrointestinal digestion

FEBS Journal ◽  
2004 ◽  
Vol 272 (2) ◽  
pp. 341-352 ◽  
Author(s):  
F. Javier Moreno ◽  
Fred A. Mellon ◽  
Martin S. J. Wickham ◽  
Andrew R. Bottrill ◽  
E. N. Clare Mills
Keyword(s):  
Major Allergen ◽  
Brazil Nut ◽  
Gastrointestinal Digestion ◽  
2S Albumin ◽  
In Vitro Gastrointestinal Digestion

scholarly journals Expression and Differential Intracellular Localization of Two Major Forms of Human 8-Oxoguanine DNA Glycosylase Encoded by Alternatively Spliced OGG1 mRNAs

1999 ◽  
Vol 10 (5) ◽  
pp. 1637-1652 ◽  
Author(s):  
Kenichi Nishioka ◽  
Toshio Ohtsubo ◽  
Hisanobu Oda ◽  
Toshiyuki Fujiwara ◽  
Dongchon Kang ◽  
...  
Keyword(s):  
Intracellular Localization ◽  
Nuclear Extract ◽  
Dna Glycosylase ◽  
Mitochondrial Targeting ◽  
Electron Microscopic ◽  
C Terminus ◽  
Alternatively Spliced ◽  
Targeting Signal ◽  
Processed Form

We identified seven alternatively spliced forms of human 8-oxoguanine DNA glycosylase (OGG1) mRNAs, classified into two types based on their last exons (type 1 with exon 7: 1a and 1b; type 2 with exon 8: 2a to 2e). Types 1a and 2a mRNAs are major in human tissues. Seven mRNAs are expected to encode different polypeptides (OGG1–1a to 2e) that share their N terminus with the common mitochondrial targeting signal, and each possesses a unique C terminus. A 36-kDa polypeptide, corresponding to OGG1–1a recognized only by antibodies against the region containing helix-hairpin-helix-PVD motif, was copurified from the nuclear extract with an activity introducing a nick into DNA containing 8-oxoguanine. A 40-kDa polypeptide corresponding to a processed form of OGG1–2a was detected in their mitochondria using antibodies against its C terminus. Electron microscopic immunocytochemistry and subfractionation of the mitochondria revealed that OGG1–2a locates on the inner membrane of mitochondria. Deletion mutant analyses revealed that the unique C terminus of OGG1–2a and its mitochondrial targeting signal are essential for mitochondrial localization and that nuclear localization of OGG1–1a depends on the NLS at its C terminus.

Reversible Denaturation of Brazil Nut 2S Albumin (Ber e1) and Implication of Structural Destabilization on Digestion by Pepsin

2005 ◽  
Vol 53 (1) ◽  
pp. 123-131 ◽  
Author(s):  
Stef J. Koppelman ◽  
Willem F. Nieuwenhuizen ◽  
Marco Gaspari ◽  
Leon M. J. Knippels ◽  
André H. Penninks ◽  
...  
Keyword(s):  
Brazil Nut ◽  
2S Albumin

scholarly journals Transketolase from Leishmania mexicana has a dual subcellular localization

2004 ◽  
Vol 382 (2) ◽  
pp. 759-767 ◽  
Author(s):  
Nicola J. VEITCH ◽  
Dante A. MAUGERI ◽  
Juan Jose CAZZULO ◽  
Ylva LINDQVIST ◽  
Michael P. BARRETT
Keyword(s):  
Subcellular Localization ◽  
Three Dimensional ◽  
Total Activity ◽  
Pentose Phosphate ◽  
Dimensional Structure ◽  
Leishmania Mexicana ◽  
Gene Encoding ◽  
X Ray Crystallography ◽  
C Terminus ◽  
Targeting Signal

Transketolase has been characterized in Leishmania mexicana. A gene encoding this enzyme was identified and cloned. The gene was expressed in Escherichia coli and the protein was purified and characterized. An apparent Km of 2.75 mM for ribose 5-phosphate was determined. X-ray crystallography was used to determine the three-dimensional structure of the enzyme to a resolution of 2.2 Å (1 Å≡0.1 nm). The C-terminus of the protein contains a type-1 peroxisome-targeting signal, suggestive of a possible glycosomal subcellular localization. Subcellular localization experiments performed with promastigote forms of the parasite revealed that the protein was predominantly cytosolic, although a significant component of the total activity was associated with the glycosomes. Transketolase is thus the first enzyme of the nonoxidative branch of the pentose phosphate pathway whose presence has been demonstrated in a peroxisome-like organelle.

scholarly journals Yeast carboxypeptidase Y vacuolar targeting signal is defined by four propeptide amino acids.

1990 ◽  
Vol 111 (2) ◽  
pp. 361-368 ◽  
Author(s):  
L A Valls ◽  
J R Winther ◽  
T H Stevens
Keyword(s):  
Amino Acids ◽  
Protein Targeting ◽  
Carboxypeptidase Y ◽  
Wild Type ◽  
Amino Terminal ◽  
Type Molecule ◽  
Amino Acid Stretch ◽  
Targeting Signal ◽  
Vacuolar Protein ◽  
Vacuolar Targeting

The amino-terminal propeptide of carboxypeptidase Y (CPY) is necessary and sufficient for targeting this glycoprotein to the vacuole of Saccharomyces cerevisiae. A 16 amino acid stretch of the propeptide was subjected to region-directed mutagenesis using randomized oligonucleotides. Mutations altering any of four contiguous amino acids, Gln-Arg-Pro-Leu, resulted in secretion of the encoded CPY precursor (proCPY), demonstrating that these residues form the core of the vacuolar targeting signal. Cells that simultaneously synthesize both wild-type and sorting-defective forms of proCPY efficiently sort and deliver only the wild-type molecule to the vacuole. These results indicate that the PRC1 missorting mutations are cis-dominant, implying that the mutant forms of proCPY are secreted as a consequence of failing to interact with the sorting apparatus, rather than a general poisoning of the vacuolar protein targeting system.

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