The Heat Treatment of P/M Tool Steels

Kulmburg, A.; Hribernik, Bruno

doi:10.4028/www.scientific.net/msf.102-104.31

Cited by 3 publications

(9 citation statements)

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“…A binding was also observed with the 10-mer by gel retardation assay but at high concentration of protein, this result is already documented in previous studies in which the size of the oligonucleotide is one of the important parameters for AlcR-DNA complex stability in gel band shift experiments [5,10].…”

Section: Binding Of Alcr(l-60) To a Specific Half-sitesupporting

confidence: 81%

“…The linker region of the protein is GST-GlySer-AlcR and three residues were added at the C-terminus. Protein expression in LCI37 Escherichia coli strain and purification were as described [10,14]. The AlcR peptide was separated from the GST by a cleavage with thrombin (Sigma) (1 U/mg of fusion protein) in presence of CaCi2 2.5 mM for 10 min at 30°C.…”

Section: Dna-binding Testmentioning

confidence: 99%

“…Electrophoretic mobility shift assays (EMSAs) were performed with a MiniProteanll apparatus from Bio-Rad. The DNA-binding shift assay was as described by Kulmburg et al [10] modified as follows: 8% polyacrylamide (30/0.36) gels were run in TBE 0.25X (24.5 mM Tris pH 8.0, 16.15 mM boric acid, 0.62 mM EDTA pH 8.0) at 4°C and 18V/cm for 30 min. A double-stranded oligonucleotide (25-mer) (see legend of Fig.…”

Section: Dna-binding Testmentioning

confidence: 99%

“…In order to approach these singularities of AlcR at the atomic level, we initiated a NMR study of the interaction of an AlcR peptide comprising the Zn-cluster with an oligonucleotide bearing the 5'-TGCGG-3' quintet. This peptide, containing the first 60 amino acids of the AlcR protein, was previously used for the identification of AlcR-specific binding sites as a fusion protein GST-AlcR(l-60) [10,11]. In this work, we demonstrate the ability of the AlcR DNA-binding domain to bind to a specific half-site DNA using gel retardation assay and NMR spectroscopy.…”

Section: Introductionmentioning

confidence: 99%

“…Fax: (33) 1-69-07-72-47. E-mail: guittet@icsn.cnrs-gif.fr Abbreviations: AlcR, alcohol regulator of Aspergillus nidulans; NMR, nuclear magnetic resonance; UAS a i c , upstream activating sequence; GST, glutathione-S-transferase; TBE, Tris-boric acid-EDTA; HPLC, high-performance liquid chromatography ; NOESY, nuclear Overhauser enhancement spectroscopy ; TOCSY, total correlation spectroscopy; E-COSY, edited correlation spectroscopy; TSP, trimethylsilyl proponiate; TPP1, time proportional phase incrementation; FID, free induction decay in the promoters of the inducible genes such as alcR [10], alcA [11], alcS and alcM [3,12]. These targets are functional and are defined as UASa/c which are responsible for the transcriptional activation by AlcR of the alcA gene [11] and of the alcR gene itself [13].…”

Section: Introductionmentioning

confidence: 99%

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The Aspergillus nidulans transcription factor AlcR forms a stable complex with its half‐site DNA: a NMR study

Cerdan¹,

Collin²,

Lenouvel

et al. 1997

FEBS Letters

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The Aspergillus nidulans transcription factor AlcR is shown by NMR and gel retardation assay to form a stable complex with oligonucleotide sequences comprising the consensus half-site 5'-TGCGG-3'. Apparent /jM dissociation constants are evaluated by both methods. The measured lifetime of the complex is 74 ±7 ms at 20°C with the following DNA sequence: 5'-C1G2T3G4C5G6G7A8T9C10-3'. The major chemical shift variations upon binding involve both the two adjacent GC pairs (G6 and G7) and, clearly, the AT pairs at both ends of the consensus sequence (T3 and A8), suggesting additional contacts of the protein with the DNA. This extensive and strong interaction with the half-site is another example of the variability in contacts of the fungal DNA-binding proteins containing Zn 2 Cys 6 domains with their consensus sites. It is the first demonstration that a binuclear cluster protein can bind to DNA as a monomer with strong affinity.

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Section: Binding Of Alcr(l-60) To a Specific Half-sitesupporting

confidence: 81%

Section: Dna-binding Testmentioning

confidence: 99%

Section: Dna-binding Testmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The Aspergillus nidulans transcription factor AlcR forms a stable complex with its half‐site DNA: a NMR study

Cerdan¹,

Collin²,

Lenouvel

et al. 1997

FEBS Letters

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Materials characterization and mechanical properties of graded tool steels processed by a new co‐spray forming technique

Cui

Schulz

Uhlenwinkel

2014

Materialwissenschaft Werkst

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In order to meet the requirements of micro cold forming tools, a new co-spray forming process has been applied to produce graded materials from two different tool steels in this study. The two steel melts were atomized and co-sprayed simultaneously onto a flat substrate, resulting in a flat graded deposit when the two sprays were overlapped. To eliminate porosity and break up carbide network, the graded deposits were further hot rolled. The resultant graded tool steels were investigated with respect to porosity, element distribution, microstructure, hardness, strength, and toughness. The degree of overlapping of the two sprays determined the concentration gradient of the chemical elements in the deposits. The overlapping of the spray cones also contributed to low porosity in the gradient zone of the deposits. The porosity in the graded deposits could be essentially eliminated by means of hot rolling. The carbides and grain structures of the hot rolled tool steels were fine and homogeneous. By means of combining different tool steels in a single deposit, different microstructures and properties were combined.

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Purification and regulation of the synthesis of a β-xylosidase from Aspergillus nidulans

Kumar¹

1996

FEMS Microbiology Letters

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β‐xylosidase (EC 3.2.1.37) has been purified from Aspergillus nidulans mycelium grown on oat‐spelt xylan as sole carbon source. Its pH optimum for activity was found to be 5.0 and the optimum temperature was 50 °C. Its molecular mass was estimated by gel filtration to be 180000. Using p‐nitrophenyl‐β‐d‐xylopyranoside as substrate, the Km and Vmax values have been found to be 1.1 mM and 25.6 μmol min−1(mg protein)−1, respectively. Enzyme activity was inhibited by Hg2+, Ag2+, and Cu2+ at a concentration of 1 × 10−3 M. The synthesis of β‐xylosidase in A. nidulans is strongly induced by arabinose and xylose and is subject to carbon catabolite repression mediated by the cre A gene product.

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The Heat Treatment of P/M Tool Steels

Cited by 3 publications

References 0 publications

The Aspergillus nidulans transcription factor AlcR forms a stable complex with its half‐site DNA: a NMR study

The Aspergillus nidulans transcription factor AlcR forms a stable complex with its half‐site DNA: a NMR study

Materials characterization and mechanical properties of graded tool steels processed by a new co‐spray forming technique

Purification and regulation of the synthesis of a β-xylosidase from Aspergillus nidulans

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