A functional characterization of the membrane bound iron sulphur centres of Rhodopseudomonas capsulata

1983 ◽  
Vol 135 (3) ◽  
pp. 176-181 ◽  
Author(s):  
Davide Zannoni ◽  
W. John Ingledew
Keyword(s):  
Functional Characterization ◽  
Rhodopseudomonas Capsulata ◽  
Membrane Bound

Isolation and Partial Characterization of the Membrane-Bound NADH Dehydrogenase from the Phototrophic Bacterium Rhodopseudomonas capsulata

1981 ◽  
Vol 36 (5-6) ◽  
pp. 400-406 ◽  
Author(s):  
Toshihisa Ohshima ◽  
Gerhart Drews
Keyword(s):  
Nadh Dehydrogenase ◽  
Deae Cellulose ◽  
Sodium Deoxycholate ◽  
Pyridine Nucleotide ◽  
Sodium Cholate ◽  
Rhodopseudomonas Capsulata ◽  
Michaelis Constant ◽  
Membrane Bound ◽  
Ph Dependent

Abstract Chemotrophically grown cells of Rhodopseudomonas capsulata contain at least three different pyridine nucleotide dehydrogenases, i) a soluble, found in the supernatant (144000 × g) of cell free extracts, NADH-dependent, ii) a mem brane-bound, NADH-dependent, and iii) a soluble, found in the supernatant N AD PH dependent. The membrane-bound NADH dehydrogenase (E.C. 1.6.99.3) has been solubilized by sodium deoxycholate treatm ent of m em branes and purified 75 fold by column chrom atography on Sephadex G-150 and DEAE cellulose in the presence of sodium cholate. The native enzyme has an apparent molecular mass (Mr) of 97 000, containing polypeptides of Mr of about 15 000. The pH optim um was at 7.5. The enzyme was specific for NADH. The Michaelis constant for NADH and DCIP were 4.0 and 63 μm, respectively. The enzyme was inactivated by FMN, riboflavin and NADH. In contrast, the soluble NADH-dehydrogenase (i) was activated by FMN.

scholarly journals Functional Characterization of the IlpA Protein of Vibrio vulnificus as an Adhesin and Its Role in Bacterial Pathogenesis

2010 ◽  
Vol 78 (6) ◽  
pp. 2408-2417 ◽  
Author(s):  
Kyung-Jo Lee ◽  
Na Yeon Lee ◽  
Yang-Soo Han ◽  
Juri Kim ◽  
Kyu-Ho Lee ◽  
...  
Keyword(s):  
Adhesion Molecule ◽  
Vibrio Vulnificus ◽  
Polyclonal Antibodies ◽  
Functional Characterization ◽  
Intestinal Epithelial ◽  
Human Intestinal Epithelial Cells ◽  
Direct Binding ◽  
Membrane Bound

ABSTRACT Vibrio vulnificus is a Gram-negative bacterium that causes a fatal septicemia. One of its virulence factors is a membrane-bound lipoprotein, IlpA, which can induce cytokine production in human immune cells. In the present study, the role of IlpA as an adhesion molecule was investigated. An ilpA-deleted V. vulnificus mutant showed significantly decreased adherence to INT-407 human intestinal epithelial cells, which in turn resulted in reduced cytotoxicity. The ΔilpA mutant recovered the adherence ability of the wild type by complementation in trans with the intact ilpA gene. In addition, pretreatment of V. vulnificus with anti-IlpA polyclonal antibodies resulted in a significant reduction of bacterial adherence. To localize the domain of IlpA required for cytoadherence, three truncated recombinant IlpA polypeptides were constructed and tested for the ability to adhere to human cells by a ligand-binding immunoblot assay and fluorescence microscopy. The polypeptide containing the carboxy (C)-terminal hydrophilic domain exhibited direct binding to INT-407 cells. Therefore, the C-terminal domain of IlpA allows this protein to be an adhesion molecule of V. vulnificus.

scholarly journals Functional Characterization of Colon-Cancer-Associated Variants in ADAM17 Affecting the Catalytic Domain

Biomedicines ◽  
2020 ◽  
Vol 8 (11) ◽  
pp. 463
Author(s):  
Jan Philipp Dobert ◽  
Anne-Sophie Cabron ◽  
Philipp Arnold ◽  
Egor Pavlenko ◽  
Stefan Rose-John ◽  
...  
Keyword(s):  
Catalytic Domain ◽  
Functional Characterization ◽  
Tumor Development ◽  
Point Mutations ◽  
Modern World ◽  
Protective Effects ◽  
Tissue Samples ◽  
Membrane Bound

Although extensively investigated, cancer is still one of the most devastating and lethal diseases in the modern world. Among different types, colorectal cancer (CRC) is most prevalent and mortal, making it an important subject of research. The metalloprotease ADAM17 has been implicated in the development of CRC due to its involvement in signaling pathways related to inflammation and cell proliferation. ADAM17 is capable of releasing membrane-bound proteins from the cell surface in a process called shedding. A deficiency of ADAM17 activity has been previously shown to have protective effects against CRC in mice, while an upregulation of ADAM17 activity is suspected to facilitate tumor development. In this study, we characterize ADAM17 variants found in tissue samples of cancer patients in overexpression studies. We here focus on point mutations identified within the catalytic domain of ADAM17 and could show a functional dysregulation of the CRC-associated variants. Since the catalytic domain of ADAM17 is the only region structurally determined by crystallography, we study the effect of each point mutation not only to learn more about the role of ADAM17 in cancer, but also to investigate the structure–function relationships of the metalloprotease.

Mechanism and Inhibition of Matrix Metalloproteinases

2019 ◽  
Vol 26 (15) ◽  
pp. 2609-2633 ◽  
Author(s):  
Linda Cerofolini ◽  
Marco Fragai ◽  
Claudio Luchinat
Keyword(s):  
Matrix Metalloproteinases ◽  
Functional Characterization ◽  
Structural Data ◽  
Therapeutic Strategies ◽  
Outer Cell ◽  
Drug Candidates ◽  
The Family ◽  
Outer Cell Membrane ◽  
Membrane Bound

Matrix metalloproteinases hydrolyze proteins and glycoproteins forming the extracellular matrix, cytokines and growth factors released in the extracellular space, and membrane-bound receptors on the outer cell membrane. The pathological relevance of MMPs has prompted the structural and functional characterization of these enzymes and the development of synthetic inhibitors as possible drug candidates. Recent studies have provided a better understanding of the substrate preference of the different members of the family, and structural data on the mechanism by which these enzymes hydrolyze the substrates. Here, we report the recent advancements in the understanding of the mechanism of collagenolysis and elastolysis, and we discuss the perspectives of new therapeutic strategies for targeting MMPs.

Functional characterization of tomato membrane-bound NAC transcription factors

2016 ◽  
Vol 93 (4-5) ◽  
pp. 511-532 ◽  
Author(s):  
Payel Bhattacharjee ◽  
Rohit Das ◽  
Arunava Mandal ◽  
Pallob Kundu
Keyword(s):  
Transcription Factors ◽  
Functional Characterization ◽  
Nac Transcription Factors ◽  
Membrane Bound

Functional characterization of soluble and membrane-bound forms of vaccinia virus complement control protein (VCP)

1999 ◽  
Vol 36 (10) ◽  
pp. 685-697 ◽  
Author(s):  
Ariella M Rosengard ◽  
Laura C Alonso ◽  
Laura C Korb ◽  
William M Baldwin ◽  
Fred Sanfilippo ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Functional Characterization ◽  
Complement Control Protein ◽  
Membrane Bound ◽  
Control Protein

scholarly journals A novel Delta12-fatty acid desaturase gene from methylotrophic yeast Pichia pastoris GS115.

2006 ◽  
Vol 53 (4) ◽  
pp. 753-759 ◽  
Author(s):  
D Sh Wei ◽  
M Ch Li ◽  
X X Zhang ◽  
H Zhou ◽  
L J Xing
Keyword(s):  
Fatty Acid ◽  
Functional Characterization ◽  
Fatty Acid Desaturase ◽  
Methylotrophic Yeast ◽  
Pastoris Gs115 ◽  
Desaturase Gene ◽  
Methylotrophic Yeast Pichia Pastoris ◽  
Membrane Bound ◽  
Fatty Acid Desaturase Gene

The methylotrophic yeast Pichia pastoris GS115, a widely used strain in production of various heterologous proteins, especially membrane-bound enzymes, can also produce linoleic and linolenic acids, which indicates the existence of membrane-bound Delta12 and Delta15-fatty acid desaturases. This paper describes the cloning and functional characterization of a novel Delta12-fatty acid desaturase gene from this methylotrophic yeast. The open reading frame of the gene (named Pp-FAD12) is 1263 bp in size and encodes a 420-amino-acid peptide. The deduced Pp-FAD12 protein shows high identity (50-67%) with Delta12-fatty acid desaturases from other fungi. It also shows a high identity (57%) with Delta15-fatty acid desaturase (named Sk-FAD15) from Saccharomyces kluyveri. Expression of Pp-FAD12 in polyunsaturated fatty acids non-producing yeast Saccharomyces cerevisiae demonstrated that its product converted oleic acid (18 : 1) to linoleic acid (18 : 2). This result suggests that Pp-FAD12 encodes a novel Delta12-fatty acid desaturase in P. pastoris GS115. This is the first report about the cloning and functional characterization of Delta12-fatty acid desaturase gene in methylotrophic yeast.

Cloning and functional characterization of a membrane-bound proinflamatory protein in Helicobacter pylori (HP-MBP1)

1998 ◽  
Vol 114 ◽  
pp. A342
Author(s):  
Masaru Yoshida ◽  
Yoshinao Kobayashi ◽  
Toshiyuki Itoh ◽  
Kazuhisa Murakami ◽  
Akira Mizoguchi ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Functional Characterization ◽  
Membrane Bound

Purification and Partial Characterization of the Soluble NADH Dehydrogenase from the Phototrophic Bacterium Rhodopseudomonas capsulata

1984 ◽  
Vol 39 (1-2) ◽  
pp. 68-72 ◽  
Author(s):  
Toshihisa Ohshima ◽  
Matsumi Ohshima ◽  
Gerhart Drews
Keyword(s):  
Cytochrome C ◽  
Nadh Dehydrogenase ◽  
Rhodopseudomonas Capsulata ◽  
Subunit Structure ◽  
Ph Optimum ◽  
Single Polypeptide Chain ◽  
Membrane Bound ◽  
Single Polypeptide ◽  
Sepharose 4B

Abstract Soluble NADH dehydrogenase was purified to homogeneity from chemotrophically grown cells of Rhodopseudomonas capsulata by ammonium sulfate fractionation, AH -Sepharose 4B chromatography and FMN-Sepharose 6B affinity chromatography. The enzyme contains a single polypeptide chain of an apparent M, of 37000, suggesting that the subunit structure is different from that of the membrane-bound enzyme. The purified soluble NADH dehydrogenase requires flavin compounds, e.g., FMN, FAD and riboflavin, for activity. Addition of FMN and FAD. but not riboflavin, to the enzyme solution stabilized the enzyme. The pH optimum for activity was at 7.5. The enzyme was specific for NADH as an electron donor while NADPH was inert. Menadione, ferricyanide, cytochrome c and DCIP served as an electron acceptor. The M ichaelis constants for NADH. DCIP, FM N. and cytochrome c were 45, 2.9. 7.9 and 15 μM, respectively. Many properties of soluble NADH dehydrogenase were substantially different from those of the membrane-bound enzyme, suggesting different functions.

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