Identification and characterization of a cDNA encoding mouse CAP: a homolog of the yeast adenylyl cyclase associated protein

CAP, an adenylyl cyclase associated protein, is present in Saccharomyces cerevisiae and Schizosaccharomyces pombe. In both organisms, CAP is bifunctional: the N-terminal domain binds to adenylyl cyclase, thereby enabling adenylyl cyclase to respond appropriately to upstream regulatory signals, such as RAS in S. cerevisiae; the C-terminal domain is required for cellular morphogenesis. Here, we describe the isolation of a cDNA encoding a CAP homolog from a higher eukaryote. The mouse CAP cDNA contains an open reading frame capable of encoding a 474 amino acid protein. The protein encoded by the mouse CAP cDNA shows extensive homology to the yeast CAP proteins, particularly in the central poly-proline rich region and in the C-terminal domain. By northern analysis, the CAP message appears to be ubiquitous, but not uniform. By indirect immunofluorescence, ectopically expressed mouse CAP protein is found in the cytoplasm of fibroblasts and, in migrating cells, at the leading edge. Expression of the mouse CAP cDNA in S. cerevisiae complements defects associated with loss of the yeast CAP carboxy-terminal domain. Hence, the function of the CAP carboxy-terminal domain has been conserved from yeast to mouse.

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Cloning and characterization of a 2,3-oxidosqualene cyclase from Eleutherococcus trifoliatus

Holzforschung ◽

10.1515/hf-2012-0120 ◽

2013 ◽

Vol 67 (4) ◽

pp. 463-471 ◽

Cited By ~ 1

Author(s):

Li-Ting Ma ◽

Sheng-Yang Wang ◽

Yen-Hsueh Tseng ◽

Yi-Ru Lee ◽

Fang-Hua Chu

Keyword(s):

Physiological Role ◽

Active Site Residues ◽

Reading Frame ◽

Oxidosqualene Cyclases ◽

Oxidosqualene Cyclase ◽

Acid Protein ◽

Leaf Tissues ◽

New Perspective ◽

Amino Acid Protein

Abstract The 2,3-oxidosqualene cyclases (OSCs) are a family of enzymes that have an important role in plant triterpene biosynthesis. In this study, an OSC gene designed EtLUS from Eleutherococcus trifoliatus has been cloned. EtLUS includes a 2292-bp open reading frame and encodes a 763-amino acid protein. EtLUS has an MLCYCR motif, which is conserved in lupeol synthases. Comparison of active-site residues and gene expression in yeast showed that EtLUS synthesizes lupeol. However, EtLUS has the highest sequence identity with β-amyrin synthases from Araliaceae rather than lupeol synthases, adding new perspective to the evolution of the OSCs of Araliaceae. Furthermore, EtLUS is upregulated in leaf tissues under methyl jasmonate treatment, which can be interpreted that lupeol and its derivatives play an ecological and physiological role in plant defense against pathogens and insect herbivores.

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Papain Does Not Cleave Operator-Bound Lambda Repressor: Structural Characterization of the Carboxy Terminal Domain and the Hinge

Journal of Biomolecular Structure and Dynamics ◽

10.1080/07391102.2001.10506688 ◽

2001 ◽

Vol 18 (4) ◽

pp. 557-567 ◽

Cited By ~ 3

Author(s):

Kaushik Ghosh ◽

Rajagopal Chattopadhyaya

Keyword(s):

Structural Characterization ◽

Lambda Repressor ◽

Terminal Domain ◽

Carboxy Terminal Domain ◽

Carboxy Terminal

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Molecular Cloning and Characterization of a cDNA Encoding Sedoheptulose-1,7-bisphosphatase from Mulberry (Morus alba var. multicaulis)

Silvae Genetica ◽

10.1515/sg-2008-0023 ◽

2008 ◽

Vol 57 (1-6) ◽

pp. 152-157 ◽

Cited By ~ 3

Author(s):

X. Ji ◽

Y. Gai ◽

J. Ma ◽

C. Zheng ◽

Z. Mu

Keyword(s):

Morus Alba ◽

Evolutionary Analysis ◽

Comparison Analysis ◽

Reading Frame ◽

Acid Protein ◽

Fructose 1,6 Bisphosphatase ◽

Rapid Amplification ◽

Molecular Evolutionary Analysis ◽

Amino Acid Protein

Abstract A full-length cDNA encoding sedoheptulose-1,7-bisphosphatase (SBPase; EC 3.1.3.37) was cloned from mulberry (Morus alba var. multicaulis) by rapid amplification of cDNA ends (RACE). The cDNA consisted of 1,527 nucleotides with an open reading frame (ORF) of 1,179 nucleotides encoding a 393 amino acid protein of approximately 42.6 kDa. Sequence comparison analysis showed that mulberry SBPase (MSBPase) had high homology to other plant counterparts. Phylogenetic and molecular evolutionary analysis revealed that MSBPase fell into plant SBPase group. Moreover, SBPase and fructose-1,6-bisphosphatase (FBPase; EC 3.1.3.11) shared 28-32% identical residues, suggesting that the two enzymes originated from the same evolution branch. Molecular modeling indicated that each subunit of MSBPase was composed of α-helices and β-sheets joined by turns and loops, and folded into a structure of hexahedron shape which was very similar to FBPase.

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NOP3 is an essential yeast protein which is required for pre-rRNA processing.

The Journal of Cell Biology ◽

10.1083/jcb.119.4.737 ◽

1992 ◽

Vol 119 (4) ◽

pp. 737-747 ◽

Cited By ~ 57

Author(s):

I D Russell ◽

D Tollervey

Keyword(s):

Northern Analysis ◽

Rna Recognition Motif ◽

Rna Recognition ◽

Rrna Processing ◽

Reading Frame ◽

Repeat Units ◽

Nucleolar Proteins ◽

Acid Protein ◽

Sds Page ◽

Amino Acid Protein

The four nucleolar proteins NOP1, SSB1, GAR1, and NSR1 of Saccharomyces cerevisiae share a repetitive domain composed of repeat units rich in glycine and arginine (GAR domain). We have cloned and sequenced a fifth member of this family, NOP3, and shown it to be essential for cell viability. The NOP3 open reading frame encodes a 415 amino acid protein with a predicted molecular mass of 45 kD, containing a GAR domain and an RNA recognition motif. NOP3-specific antibodies recognize a 60-kD protein by SDS-PAGE and decorate the nucleolus and the surrounding nucleoplasm. A conditional lethal mutation, GAL::nop3, was constructed; growth of the mutant strain in glucose medium represses NOP3 expression. In cells depleted of NOP3, production of cytoplasmic ribosomes is impaired. Northern analysis and pulse-chase labeling indicate that pre-rRNA processing is inhibited at the late steps, in which 27SB pre-rRNA is cleaved to 25S rRNA and 20S pre-rRNA to 18S rRNA.

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Functional and structural characterization of the kinase insert and the carboxy terminal domain in VEGF receptor 2 activation

The FASEB Journal ◽

10.1096/fj.14-256206 ◽

2014 ◽

Vol 28 (11) ◽

pp. 4914-4923 ◽

Cited By ~ 8

Author(s):

Sandro Manni ◽

Kaisa Kisko ◽

Thomas Schleier ◽

Jack Missimer ◽

Kurt Ballmer‐Hofer

Keyword(s):

Structural Characterization ◽

Vegf Receptor ◽

Terminal Domain ◽

Carboxy Terminal Domain ◽

Carboxy Terminal

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Construction and Characterization of Chimeric Enzyme Consisting of an Amino-Terminal Domain of Phenylalanine Dehydrogenase and a Carboxy-Terminal Domain of Leucine Dehydrogenase

The Journal of Biochemistry ◽

10.1093/oxfordjournals.jbchem.a124618 ◽

1994 ◽

Vol 116 (4) ◽

pp. 931-936 ◽

Cited By ~ 11

Author(s):

Kunishige Kataoka ◽

Harumi Takada ◽

Katsuyuki Tanizawa ◽

Tohru Yoshimura ◽

Nobuyoshi Esaki ◽

...

Keyword(s):

Chimeric Enzyme ◽

Phenylalanine Dehydrogenase ◽

Amino Terminal ◽

Leucine Dehydrogenase ◽

Terminal Domain ◽

Carboxy Terminal Domain ◽

Carboxy Terminal ◽

Amino Terminal Domain

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Characterization of an Endo-β-1,6-Galactanase from Streptomyces avermitilis NBRC14893

Applied and Environmental Microbiology ◽

10.1128/aem.01733-07 ◽

2008 ◽

Vol 74 (8) ◽

pp. 2379-2383 ◽

Cited By ~ 16

Author(s):

Hitomi Ichinose ◽

Toshihisa Kotake ◽

Yoichi Tsumuraya ◽

Satoshi Kaneko

Keyword(s):

Signal Sequence ◽

Recombinant Enzyme ◽

Streptomyces Avermitilis ◽

Glycoside Hydrolase Family ◽

Reading Frame ◽

Secretion Signal ◽

Acid Protein ◽

Hydrolysis Of ◽

Amino Acid Protein

ABSTRACT The putative endo-β-1,6-galactanase gene from Streptomyces avermitilis was cloned and expressed in Escherichia coli, and the enzymatic properties of the recombinant enzyme were characterized. The gene consisted of a 1,476-bp open reading frame and encoded a 491-amino-acid protein, comprising an N-terminal secretion signal sequence and glycoside hydrolase family 5 catalytic module. The recombinant enzyme, Sa1,6Gal5A, catalyzed the hydrolysis of β-1,6-linked galactosyl linkages of oligosaccharides and polysaccharides. The enzyme produced galactose and a range of β-1,6-linked galacto-oligosaccharides, predominantly β-1,6-galactobiose, from β-1,6-galactan chains. There was a synergistic effect between the enzyme and Sa1,3Gal43A in degrading tomato arabinogalactan proteins. These results suggest that Sa1,6Gal5A is the first identified endo-β-1,6-galactanase from a prokaryote.

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Molecular Cloning and Biochemical Characterization of a Recombinant Sterol 3-O-Glucosyltransferase fromGymnema sylvestreR.Br. Catalyzing Biosynthesis of Steryl Glucosides

BioMed Research International ◽

10.1155/2014/934351 ◽

2014 ◽

Vol 2014 ◽

pp. 1-14 ◽

Cited By ~ 10

Author(s):

Pragya Tiwari ◽

Rajender Singh Sangwan ◽

Asha ◽

B. N. Mishra ◽

Farzana Sabir ◽

...

Keyword(s):

Biochemical Characterization ◽

Present Report ◽

Hydroxyl Group ◽

Biological Origin ◽

Reading Frame ◽

Steryl Glucosides ◽

Acid Protein ◽

Phytochemical Profiles ◽

Amino Acid Protein

Gymnema sylvestreR.Br., a pharmacologically important herb vernacularly called Gur-Mar (sugar eliminator), is widely known for its antidiabetic action. This property of the herb has been attributed to the presence of bioactive triterpene glycosides. Although some information regarding pharmacology and phytochemical profiles of the plant are available, no attempts have been made so far to decipher the biosynthetic pathway and key enzymes involved in biosynthesis of steryl glucosides. The present report deals with the identification and catalytic characterization of a glucosyltransferase, catalyzing biosynthesis of steryl glycosides. The full length cDNA (2572 bp) contained an open reading frame of 2106 nucleotides that encoded a 701 amino acid protein, falling into GT-B subfamily of glycosyltransferases. The GsSGT was expressed inEscherichia coliand biochemical characterization of the recombinant enzyme suggested its key role in the biosynthesis of steryl glucosides with catalytic preference for C-3 hydroxyl group of sterols. To our knowledge, this pertains to be the first report on cloning and biochemical characterization of a sterol metabolism gene fromG. sylvestreR.Br. catalyzing glucosylation of a variety of sterols of biological origin from diverse organisms such as bacteria, fungi, and plants.

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Characterization of a Phosphoglucose Isomerase-like Activity Associated with the Carboxy-Terminal Domain ofEscherichia coliGlucosamine-6-phosphate Synthase

Journal of the American Chemical Society ◽

10.1021/ja953614q ◽

1996 ◽

Vol 118 (7) ◽

pp. 1797-1798 ◽

Cited By ~ 25

Author(s):

Caroline Leriche ◽

Marie-Ange Badet-Denisot ◽

Bernard Badet

Keyword(s):

Phosphoglucose Isomerase ◽

Terminal Domain ◽

Carboxy Terminal Domain ◽

Carboxy Terminal ◽

Phosphate Synthase

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Isolation and characterization of the muscle-specific isoform of creatine kinase from the zebrafish, Danio rerio

Biochemistry and Cell Biology ◽

10.1139/o01-134 ◽

2001 ◽

Vol 79 (6) ◽

pp. 779-782 ◽

Cited By ~ 6

Author(s):

Gregory Harder ◽

Ross McGowan

Keyword(s):

Creatine Kinase ◽

Amino Acid ◽

Danio Rerio ◽

Cdna Sequence ◽

Reading Frame ◽

Isolation And Characterization ◽

Acid Protein ◽

Amino Acid Levels ◽

Amino Acid Protein

We have isolated and characterized a cDNA sequence corresponding to the zebrafish muscle-specific isoform of creatine kinase. The sequence is 1552 bases in length and contains an open reading frame capable of producing a 381 amino acid protein. The sequence is very similar to muscle-specific creatine kinases isolated from other species at both the nucleotide and amino acid levels but contains some differences from a previously reported zebrafish clone.Key words: creatine kinase, muscle isoform, zebrafish, Danio rerio.

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