Munc13-4 is a GTP-Rab27-binding protein regulating dense core granule secretion in platelets.
AuthorsShirakawa R, Higashi T, Tabuchi A, Yoshioka A, Nishioka H, Fukuda M, Kita T, Horiuchi H,
JournalJ Biol Chem
PubMed ID14699162
'Platelets store self-agonists such as ADP and serotonin in dense core granules. Although exocytosis of these granules is crucial for hemostasis and thrombosis, the underlying mechanism is not fully understood. Here, we show that incubation of permeabilized platelets with unprenylated active mutant Rab27A-Q78L, wild type Rab27A, and Rab27B inhibited the ... More
Crystal structure of Thermotoga maritima alpha-L-fucosidase. Insights into the catalytic mechanism and the molecular basis for fucosidosis.
AuthorsSulzenbacher G, Bignon C, Nishimura T, Tarling CA, Withers SG, Henrissat B, Bourne Y,
JournalJ Biol Chem
PubMed ID14715651
'Fucosylated glycoconjugates are involved in numerous biological events, and alpha-l-fucosidases, the enzymes responsible for their processing, are therefore of crucial importance. Deficiency in alpha-l-fucosidase activity is associated with fucosidosis, a lysosomal storage disorder characterized by rapid neurodegeneration, resulting in severe mental and motor deterioration. To gain insight into alpha-l-fucosidase function ... More
The oncoprotein Tax binds the SRC-1-interacting domain of CBP/p300 to mediate transcriptional activation.
AuthorsScoggin KE, Ulloa A, Nyborg JK,
JournalMol Cell Biol
PubMed ID11463834
'Oncogenesis associated with human T-cell leukemia virus (HTLV) infection is directly linked to the virally encoded transcription factor Tax. To activate HTLV-1 transcription Tax interacts with the cellular protein CREB and the pleiotropic coactivators CBP and p300. While extensively studied, the molecular mechanisms of Tax transcription function and coactivator utilization ... More
Human DNA polymerase N (POLN) is a low fidelity enzyme capable of error-free bypass of 5S-thymine glycol.
AuthorsTakata K, Shimizu T, Iwai S, Wood RD,
JournalJ Biol Chem
PubMed ID16787914
'Human DNA polymerase N (POLN or pol nu) is the most recently discovered nuclear DNA polymerase in the human genome. It is an A-family DNA polymerase related to Escherichia coli pol I, human POLQ, and Drosophila Mus308. We report the first purification of the recombinant enzyme and examination of its ... More
Dual-tagging system for the affinity purification of mammalian protein complexes.
AuthorsGiannone RJ, McDonald WH, Hurst GB, Huang Y, Wu J, Liu Y, Wang Y,
JournalBiotechniques
PubMed ID17907572
'Although affinity purification coupled with mass spectrometry (MS) provides a powerful tool to study protein-protein interactions, this strategy has encountered numerous difficulties when adapted to mammalian cells. Here we describe a Gateway-compatible dual-tag affinity purification system that integrates regulatable expression, tetracysteine motifs, and various combinations ofaffinity tags to facilitate the ... More
Biosynthesis pathway of ADP-L-glycero-beta-D-manno-heptose in Escherichia coli.
'The steps involved in the biosynthesis of the ADP-L-glycero-beta-D-manno-heptose (ADP-L-beta-D-heptose) precursor of the inner core lipopolysaccharide (LPS) have not been completely elucidated. In this work, we have purified the enzymes involved in catalyzing the intermediate steps leading to the synthesis of ADP-D-beta-D-heptose and have biochemically characterized the reaction products by ... More
Identification of an antiangiogenic FGF2-binding site in the N terminus of the soluble pattern recognition receptor PTX3.
AuthorsCamozzi M, Rusnati M, Bugatti A, Bottazzi B, Mantovani A, Bastone A, Inforzato A, Vincenti S, Bracci L, Mastroianni D, Presta M,
JournalJ Biol Chem
PubMed ID16769728
'Long-pentraxin 3 (PTX3) is a soluble pattern recognition receptor with non-redundant functions in inflammation and innate immunity. PTX3 comprises a pentraxin-like C-terminal domain involved in complement activation via C1q interaction and an N-terminal extension with unknown functions. PTX3 binds fibroblast growth factor-2 (FGF2), inhibiting its pro-angiogenic and pro-restenotic activity. Here, ... More
Ubiquitination of a novel deubiquitinating enzyme requires direct binding to von Hippel-Lindau tumor suppressor protein.
Authors Li Zaibo; Na Xi; Wang Dakun; Schoen Susan R; Messing Edward M; Wu Guan;
JournalJ Biol Chem
PubMed ID11739384
'von Hippel-Lindau (VHL) disease is a hereditary cancer syndrome caused by germline mutations of the VHL gene. Recent studies suggest that VHL protein (pVHL) is a component of an E3 ubiquitin ligase, but the detailed biological function of pVHL remains to be determined. To further elucidate the biological functions of ... More
Modular structure of the receptor binding proteins of Lactococcus lactis phages. The RBP structure of the temperate phage TP901-1.
Lactococcus lactis is a gram-positive bacterium widely used by the dairy industry. Several industrial L. lactis strains are sensitive to various distinct bacteriophages. Most of them belong to the Siphoviridae family and comprise several species, among which the 936 and P335 are prominent. Members of these two phage species recognize ... More
Molecular and Biochemical Characterization of a Novel Intracellular Invertase from Aspergillus niger with Transfructosylating Activity.
AuthorsGoosen C, Yuan XL, van Munster JM, Ram AF, van der Maarel MJ, Dijkhuizen L,
JournalEukaryot Cell
PubMed ID17293485
A novel sub-family of putative intracellular invertase enzymes (glycoside hydrolase family 32) has previously been identified in fungal genomes. Here we report phylogenetic, molecular and biochemical characteristics of SucB, one of two novel intracellular invertases identified in Aspergillus niger. The sucB gene was expressed in Escherichia coli and an invertase ... More
Cytosolic Hydroxymethyldihydropterin Pyrophosphokinase/Dihydropteroate Synthase from Arabidopsis thaliana: A SPECIFIC ROLE IN EARLY DEVELOPMENT AND STRESS RESPONSE.
AuthorsStorozhenko S, Navarrete O, Ravanel S, De Brouwer V, Chaerle P, Zhang GF, Bastien O, Lambert W, Rébeillé F, Van Der Straeten D,
JournalJ Biol Chem
PubMed ID17289662
In plants, 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase/7,8-dihydropteroate synthase (mitHPPK/DHPS) is a bifunctional mitochondrial enzyme, which catalyzes the first two consecutive steps of tetrahydrofolate biosynthesis. Mining the Arabidopsis genome data base has revealed a second gene encoding a protein that lacks a potential transit peptide, suggesting a cytosolic localization of the isoenzyme (cytHPPK/DHPS). When ... More
A serpin from the gut bacterium Bifidobacterium longum inhibits eukaryotic elastase-like serine proteases.
AuthorsIvanov D, Emonet C, Foata F, Affolter M, Delley M, Fisseha M, Blum-Sperisen S, Kochhar S, Arigoni F,
JournalJ Biol Chem
PubMed ID16627467
Serpins form a large class of protease inhibitors involved in regulation of a wide spectrum of physiological processes. Recently identified prokaryotic members of this protein family may provide a key to the evolutionary origins of the unique serpin fold and the associated inhibitory mechanism. We performed a biochemical characterization of ... More
Expression and characterization of the protein Rv1399c from Mycobacterium tuberculosis. A novel carboxyl esterase structurally related to the HSL family.
The Mycobacterium tuberculosis genome contains an unusually high number of proteins involved in the metabolism of lipids belonging to the Lip family, including various nonlipolytic and lipolytic hydrolases. Driven by a structural genomic approach, we have biochemically characterized the Rv1399c gene product, LipH, previously annotated as a putative lipase. Rv1399c ... More
Fbx15 is a novel target of Oct3/4 but is dispensable for embryonic stem cell self-renewal and mouse development.
AuthorsTokuzawa Y, Kaiho E, Maruyama M, Takahashi K, Mitsui K, Maeda M, Niwa H, Yamanaka S,
JournalMol Cell Biol
PubMed ID12665572
Embryonic stem (ES) cells are immortal and pluripotent cells derived from early mammalian embryos. Transcription factor Oct3/4 is essential for self-renewal of ES cells and early mouse development. However, only a few Oct3/4 target genes have been identified. In this study, we found that F-box-containing protein Fbx15 was expressed predominantly ... More
Identification and characterization of the human mus81-eme1 endonuclease.
AuthorsCiccia A, Constantinou A, West SC,
JournalJ Biol Chem
PubMed ID12721304
The faithful and complete replication of DNA is necessary for the maintenance of genome stability. It is known, however, that replication forks stall at lesions in the DNA template and need to be processed so that replication restart can occur. In fission yeast, the Mus81-Eme1 endonuclease complex (Mus81-Mms4 in Saccharomyces ... More
DEN1 Is a Dual Function Protease Capable of Processing the C Terminus of Nedd8 and Deconjugating Hyper-neddylated CUL1.
AuthorsWu K, Yamoah K, Dolios G, Gan-Erdene T, Tan P, Chen A, Lee CG, Wei N, Wilkinson KD, Wang R, Pan ZQ,
JournalJ Biol Chem
PubMed ID12759363
Nedd8 activates ubiquitination by increasing the efficiency of polyubiquitin chain assembly through its covalent conjugation to cullin molecules. Here we report the isolation, cloning, and characterization of a novel human Nedd8-specific protease called DEN1. Human DEN1 is encoded by AAH31411.1, a previously uncharacterized protein of 212 amino acids that shares ... More
PAIR2 is essential for homologous chromosome synapsis in rice meiosis I.
AuthorsNonomura K, Nakano M, Eiguchi M, Suzuki T, Kurata N,
JournalJ Cell Sci
PubMed ID16410547
The PAIR2 gene is required for homologous chromosome synapsis at meiosis I in rice (Oryza sativa L.) and encodes a HORMA-domain protein that is homologous to Saccharomyces cerevisiae HOP1 and Arabidopsis ASY1. Immunocytological and electron microscopic analyses indicate that PAIR2 proteins associate with axial elements (AEs) at leptotene and zygotene, ... More
DNA cloning using in vitro site-specific recombination.
Authors Hartley J L; Temple G F; Brasch M A;
JournalGenome Res
PubMed ID11076863
As a result of numerous genome sequencing projects, large numbers of candidate open reading frames are being identified, many of which have no known function. Analysis of these genes typically involves the transfer of DNA segments into a variety of vector backgrounds for protein expression and functional analysis. We describe ... More
Formation of the approximately 350-kDa Apg12-Apg5.Apg16 multimeric complex, mediated by Apg16 oligomerization, is essential for autophagy in yeast.
Autophagy, responsible for the delivery of cytoplasmic components to the lysosome/vacuole for degradation, is the major degradative pathway in eukaryotic cells. This process requires a ubiquitin-like protein conjugation system, in which Apg12 is covalently bound to Apg5. In the yeast Saccharomyces cerevisiae, the Apg12-Apg5 conjugate further interacts with a small ... More
A cytosolic form of aminopeptidase P from Drosophila melanogaster: molecular cloning and characterization.
Authors Kulkarni Gauri V; Deobagkar Deepti D;
JournalJ Biochem (Tokyo)
PubMed ID11872174
Using a functional genomic approach, we have identified and characterized a cytosolic form of aminopeptidase P from Drosophila melanogaster. This study represents the first characterization of an insect aminopeptidase P. The complete sequence of a 12.5 kbp genomic clone from D. melanogaster showed the presence of a 1,839 bp ORF, ... More
Carbonic anhydrase II binds to and enhances activity of the Na+/H+ exchanger.
Authors Li Xiuju; Alvarez Bernardo; Casey Joseph R; Reithmeier Reinhart A F; Fliegel Larry;
JournalJ Biol Chem
PubMed ID12138085
We examined the ability of carbonic anhydrase II to bind to and affect the transport efficiency of the NHE1 isoform of the mammalian Na(+)/H(+) exchanger. The C-terminal region of NHE1 was expressed in Escherichia coli fused with an N-terminal glutathionine S-transferase or with a C-terminal polyhistidine tag. Using a microtiter ... More
Biosynthesis of nucleotide-activated D-glycero-D-manno-heptose.
Authors Kneidinger B; Graninger M; Puchberger M; Kosma P; Messner P;
JournalJ Biol Chem
PubMed ID11279237
The glycan chain repeats of the S-layer glycoprotein of Aneurinibacillus thermoaerophilus DSM 10155 contain d-glycero-d-manno-heptose, which has also been described as constituent of lipopolysaccharide cores of Gram-negative bacteria. The four genes required for biosynthesis of the nucleotide-activated form GDP-d-glycero-d-manno-heptose were cloned, sequenced, and overexpressed in Escherichia coli, and the corresponding ... More
Identification of two GDP-6-deoxy-D-lyxo-4-hexulose reductases synthesizing GDP-D-rhamnose in Aneurinibacillus thermoaerophilus L420-91T.
Authors Kneidinger B; Graninger M; Adam G; Puchberger M; Kosma P; Zayni S; Messner P;
JournalJ Biol Chem
PubMed ID11096116
The glycan repeats of the surface layer glycoprotein of Aneurinibacillus thermoaerophilus L420-91T contain d-rhamnose and 3-acetamido-3,6-dideoxy-d-galactose, both of which are also constituents of lipopolysaccharides of Gram-negative plant and human pathogenic bacteria. The two genes required for biosynthesis of the nucleotide-activated precursor GDP-d-rhamnose, gmd and rmd, were cloned, sequenced, and overexpressed ... More
Proteome-scale purification of human proteins from bacteria.
The completion of the human genome project and the development of high-throughput approaches herald a dramatic acceleration in the pace of biological research. One of the most compelling next steps will be learning the functional roles of all proteins. Achievement of this goal depends in part on the rapid expression ... More
C-terminal domain phosphatase-like family members (AtCPLs) differentially regulate Arabidopsis thaliana abiotic stress signaling, growth, and development.
Authors Koiwa Hisashi; Barb Adam W; Xiong Liming; Li Fang; McCully Michael G; Lee Byeong-Ha; Sokolchik Irina; Zhu Jianhua; Gong Zhizhong; Reddy Muppala; Sharkhuu Altanbadralt; Manabe Yuzuki; Yokoi Shuji; Zhu Jian-Kang; Bressan Ray A; Hasegawa Paul M;
JournalProc Natl Acad Sci U S A
PubMed ID12149434
Cold, hyperosmolarity, and abscisic acid (ABA) signaling induce RD29A expression, which is an indicator of the plant stress adaptation response. Two nonallelic Arabidopsis thaliana (ecotype C24) T-DNA insertional mutations, cpl1 and cpl3, were identified based on hyperinduction of RD29A expression that was monitored by using the luciferase (LUC) reporter gene ... More