scholarly journals Structural and functional characterization of the Geobacillus copper nitrite reductase: Involvement of the unique N-terminal region in the interprotein electron transfer with its redox partner

2014 ◽  
Vol 1837 (3) ◽  
pp. 396-405 ◽  
Author(s):  
Yohta Fukuda ◽  
Hiroyasu Koteishi ◽  
Ryohei Yoneda ◽  
Taro Tamada ◽  
Hideto Takami ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Nitrite Reductase ◽  
Functional Characterization ◽  
Terminal Region ◽  
Redox Partner

Heterologous production and functional characterization of Bradyrhizobium japonicum copper-containing nitrite reductase and its physiological redox partner cytochrome c550

Metallomics ◽  
2020 ◽  
Author(s):  
Julio C. Cristaldi ◽  
Felix M. Ferroni ◽  
Andrea B. Duré ◽  
Cintia S. Ramírez ◽  
Sergio D. Dalosto ◽  
...  
Keyword(s):  
Electron Donor ◽  
Nitrite Reductase ◽  
Bradyrhizobium Japonicum ◽  
Functional Characterization ◽  
Heterologous Production ◽  
Redox Partner

The copper-containing nitrite reductase from Bradyrhizobium japonicum reduces nitrite using a monoheme cytochrome c550 as an electron donor.

scholarly journals Human xylosyltransferase I and N-terminal truncated forms: functional characterization of the core enzyme

2006 ◽  
Vol 394 (1) ◽  
pp. 163-171 ◽  
Author(s):  
Sandra Müller ◽  
Jennifer Disse ◽  
Manuela Schöttler ◽  
Sylvia Schön ◽  
Christian Prante ◽  
...  
Keyword(s):  
Amino Acids ◽  
Catalytic Activity ◽  
Binding Capacity ◽  
Functional Characterization ◽  
Terminal Region ◽  
Binding Motif ◽  
Heparin Binding ◽  
Loss Of Function ◽  
The Impact

Human XT-I (xylosyltransferase I; EC 2.4.2.26) initiates the biosynthesis of the glycosaminoglycan linkage region and is a diagnostic marker of an enhanced proteoglycan biosynthesis. In the present study, we have investigated mutant enzymes of human XT-I and assessed the impact of the N-terminal region on the enzymatic activity. Soluble mutant enzymes of human XT-I with deletions at the N-terminal domain were expressed in insect cells and analysed for catalytic activity. As many as 260 amino acids could be truncated at the N-terminal region of the enzyme without affecting its catalytic activity. However, truncation of 266, 272 and 273 amino acids resulted in a 70, 90 and >98% loss in catalytic activity. Interestingly, deletion of the single 12 amino acid motif G261KEAISALSRAK272 leads to a loss-of-function XT-I mutant. This is in agreement with our findings analysing the importance of the Cys residues where we have shown that C276A mutation resulted in a nearly inactive XT-I enzyme. Moreover, we investigated the location of the heparin-binding site of human XT-I using the truncated mutants. Heparin binding was observed to be slightly altered in mutants lacking 289 or 568 amino acids, but deletion of the potential heparin-binding motif P721KKVFKI727 did not lead to a loss of heparin binding capacity. The effect of heparin or UDP on the XT-I activity of all mutants was not significantly different from that of the wild-type. Our study demonstrates that over 80% of the nucleotide sequence of the XT-I-cDNA is necessary for expressing a recombinant enzyme with full catalytic activity.

scholarly journals Expression of Pseudomonas aeruginosa nitrite reductase in Pseudomonas putida and characterization of the recombinant protein

1992 ◽  
Vol 285 (2) ◽  
pp. 661-666 ◽  
Author(s):  
M C Silvestrini ◽  
F Cutruzzolà ◽  
R D'Alessandro ◽  
M Brunori ◽  
N Fochesato ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Electron Transfer ◽  
Recombinant Protein ◽  
Pseudomonas Putida ◽  
Molecular Mass ◽  
Nitrite Reductase ◽  
Water Soluble ◽  
Binding Kinetics ◽  
Cytochrome C551

Nitrite reductase from Pseudomonas aeruginosa has been successfully expressed in Pseudomonas putida. The purified recombinant enzyme contains haem c but no haem d1. Nonetheless, like the holoenzyme from Ps. aeruginosa, it is a stable dimer (molecular mass 120 kDa), and electron transfer to oxidized azurin is biphasic and follows bimolecular kinetics (k1 = 1.5 x 10(5) and k2 = 2.2 x 10(4) M-1.s-1). Unlike the chemically produced apoenzyme, recombinant nitrite reductase containing only haem c is water-soluble, stable at neutral pH and can be quantitatively reconstituted with haem d1, yielding a holoenzyme with the same properties as that expressed by Ps. aeruginosa (namely optical and c.d. spectra, molecular mass, cytochrome c551 oxidase activity and CO-binding kinetics).

scholarly journals Functional Characterization of an Amino-terminal Region of HDAC4 That Possesses MEF2 Binding and Transcriptional Repressive Activity

2003 ◽  
Vol 278 (26) ◽  
pp. 23515-23521 ◽  
Author(s):  
Jonathan K. L. Chan ◽  
Luguo Sun ◽  
Xiang-Jiao Yang ◽  
Guang Zhu ◽  
Zhenguo Wu
Keyword(s):  
Functional Characterization ◽  
Terminal Region ◽  
Amino Terminal

Structural and functional characterization of an epitope in the conserved C-terminal region of HIV-1 gp120

1999 ◽  
Vol 54 (1) ◽  
pp. 32-42 ◽  
Author(s):  
M. Ferrer ◽  
B.J. Sullivan ◽  
K.L. Godbout ◽  
E. Burke ◽  
H.S. Stump ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Terminal Region ◽  
Hiv 1

scholarly journals Functional Characterization of the N-terminal Region of Myosin-2

2006 ◽  
Vol 281 (47) ◽  
pp. 36102-36109 ◽  
Author(s):  
Setsuko Fujita-Becker ◽  
Georgios Tsiavaliaris ◽  
Reiko Ohkura ◽  
Takashi Shimada ◽  
Dietmar J. Manstein ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Terminal Region

On the road to improve the bioremediation and electricity-harvesting skills of Geobacter sulfurreducens: functional and structural characterization of multihaem cytochromes

2012 ◽  
Vol 40 (6) ◽  
pp. 1295-1301 ◽  
Author(s):  
Leonor Morgado ◽  
Ana P. Fernandes ◽  
Joana M. Dantas ◽  
Marta A. Silva ◽  
Carlos A. Salgueiro
Keyword(s):  
Electron Transfer ◽  
Functional Characterization ◽  
Cost Effective ◽  
Genetically Engineered ◽  
Geobacter Sulfurreducens ◽  
Extracellular Electron Transfer ◽  
The Road ◽  
On The Road ◽  
Haem Proteins

Extracellular electron transfer is one of the physiological hallmarks of Geobacter sulfurreducens, allowing these bacteria to reduce toxic and/or radioactive metals and grow on electrode surfaces. Aiming to functionally optimize the respiratory electron-transfer chains, such properties can be explored through genetically engineered strains. Geobacter species comprise a large number of different multihaem c-type cytochromes involved in the extracellular electron-transfer pathways. The functional characterization of multihaem proteins is particularly complex because of the coexistence of several microstates in solution, connecting the fully reduced and oxidized states. NMR spectroscopy has been used to monitor the stepwise oxidation of each individual haem and thus to obtain information on each microstate. For the structural study of these proteins, a cost-effective isotopic labelling of the protein polypeptide chains was combined with the comparative analysis of 1H-13C HSQC (heteronuclear single-quantum correlation) NMR spectra obtained for labelled and unlabelled samples. These new methodological approaches allowed us to study G. sulfurreducens haem proteins functionally and structurally, revealing functional mechanisms and key residues involved in their electron-transfer capabilities. Such advances can now be applied to the design of engineered haem proteins to improve the bioremediation and electricity-harvesting skills of G. sulfurreducens.

scholarly journals Functional characterization of mutants deficient in N-terminal phosphorylation of the CF1 subunit beta

2021 ◽  
Author(s):  
Ralph Bock ◽  
Deserah D Strand ◽  
Daniel Karcher ◽  
Stephanie Ruf ◽  
Anne Schadach ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Electron Transport ◽  
Atp Synthase ◽  
Electron Transport Chain ◽  
Atp Hydrolysis ◽  
Molecular Motor ◽  
Functional Characterization ◽  
Phosphorylation Sites ◽  
Transport Chain

Understanding the regulation of photosynthetic light harvesting and electron transfer is of great importance to efforts to improve the ability of the electron transport chain to supply downstream metabolism. The central regulator of the electron transport chain is the ATP synthase, the molecular motor that harnesses the chemiosmotic potential generated from proton coupled electron transport to synthesize ATP. The ATP synthase is regulated both thermodynamically and post-translationally, with proposed phosphorylation sites on multiple subunits. In this study we focused on two N-terminal serines on the catalytic subunit beta, previously proposed to be important for dark inactivation of the complex to avoid ATP hydrolysis at night. Here we show that there is no clear role for phosphorylation in the dark inactivation of ATP synthase. Instead, mutation of one of the two phosphorylated serine residues to aspartate strongly decreased ATP synthase abundance. We propose that the loss of N-terminal phosphorylation of ATP beta may be involved in proper ATP synthase accumulation during complex assembly.

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