scholarly journals Post-translational modification is essential for catalytic activity of nitrile hydratase

2000 ◽  
Vol 9 (5) ◽  
pp. 1024-1030 ◽  
Author(s):  
Taku Murakami ◽  
Masaki Nojiri ◽  
Hiroshi Nakayama ◽  
Naoshi Dohmae ◽  
Koji Takio ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Nitrile Hydratase ◽  
Post Translational Modification

scholarly journals The Alpha Subunit of Nitrile Hydratase Is Sufficient for Catalytic Activity and Post-Translational Modification

Biochemistry ◽  
2014 ◽  
Vol 53 (24) ◽  
pp. 3990-3994 ◽  
Author(s):  
Micah T. Nelp ◽  
Andrei V. Astashkin ◽  
Linda A. Breci ◽  
Reid M. McCarty ◽  
Vahe Bandarian
Keyword(s):  
Catalytic Activity ◽  
Nitrile Hydratase ◽  
Alpha Subunit ◽  
Post Translational Modification

scholarly journals PAR recognition by multiple reader domains of PARP1 allosterically regulates the DNA-dependent activities and independently stimulates the catalytic activity of PARP1

2021 ◽  
Author(s):  
Waghela Deeksha ◽  
Suman Abhishek ◽  
Eerappa Rajakumara
Keyword(s):  
Catalytic Activity ◽  
Repair Process ◽  
Single Strand ◽  
Genomic Integrity ◽  
Post Translational Modification ◽  
Dna Breaks ◽  
Binding Studies ◽  
Competition Binding ◽  
Multiple Reader ◽  
Stimulation Of

Poly(ADP-ribosyl)ation is a post translational modification, predominantly catalyzed by Poly(ADP-ribose) polymerase 1 (PARP1) in response to DNA damage, mediating the DNA repair process to maintain genomic integrity. Single strand (SSB) and double strand (DSB) DNA breaks are bonafide stimulators of PARP1 activity. We identified that, in addition, single strand (ss) DNA also binds and stimulates the PARP1 activity. Poly(ADP-ribose) (PAR) is chemically similar to ssDNA. However, PAR mediated PARP1 regulation remains unexplored. Here, we report ZnF3, BRCT and WGR, hitherto uncharacterized, as PAR-specific reader domains of PARP1. Surprisingly, these domains recognize PARylated protein with a higher affinity compared to PAR, but do not bind to DNA. Conversely, N-terminal domains, ZnF1 and ZnF2, of PARP1 recognize DNA but not PAR. Further competition binding studies suggest that PAR binding, allosterically releases DNA from PARP1. Unexpectedly, PAR showed catalytic stimulation of PARP1 but hampers the DNA dependent stimulation. Altogether, our work discovers dedicated PAR and DNA reader domains of the PARP1, and uncovers a novel mechanism of allosteric stimulation of the catalytic activity of PARP1 but retardation of DNA-dependent activities of PARP1 by its catalytic product PAR.

scholarly journals Poly(ADP-ribose) glycohydrolase promotes formation and homology-directed repair of meiotic DNA double-strand breaks independent of its catalytic activity

2020 ◽  
Author(s):  
Eva Janisiw ◽  
Marilina Raices ◽  
Fabiola Balmir ◽  
Luis Paulin Paz ◽  
Antoine Baudrimont ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Synaptonemal Complex ◽  
Double Strand Breaks ◽  
Dna Double Strand Breaks ◽  
Post Translational Modification ◽  
Dna Breaks ◽  
Strand Breaks ◽  
Homology Directed Repair ◽  
Damage Repair ◽  
Somatic Tissues

SummaryPoly(ADP-ribosyl)ation is a reversible post-translational modification synthetized by ADP-ribose transferases and removed by poly(ADP-ribose) glycohydrolase (PARG), which plays important roles in DNA damage repair. While well-studied in somatic tissues, much less is known about poly(ADP-ribosyl)ation in the germline, where DNA double-strand breaks are introduced by a regulated program and repaired by crossover recombination to establish a tether between homologous chromosomes. The interaction between the parental chromosomes is facilitated by meiotic specific adaptation of the chromosome axes and cohesins, and reinforced by the synaptonemal complex. Here, we uncover an unexpected role for PARG in promoting the induction of meiotic DNA breaks and their homologous recombination-mediated repair in Caenorhabditis elegans. PARG-1/PARG interacts with both axial and central elements of the synaptonemal complex, REC-8/Rec8 and the MRN/X complex. PARG-1 shapes the recombination landscape and reinforces the tightly regulated control of crossover numbers without requiring its catalytic activity. We unravel roles in regulating meiosis, beyond its enzymatic activity in poly(ADP-ribose) catabolism.

An anchoring residue adjacent to the substrate access tunnel entrance of nitrile hydratase directs its catalytic activity towards 3-cyanopyridine

2021 ◽  
Author(s):  
Zhongyi Cheng ◽  
Weimiao Zhang ◽  
Yuanyuan Xia ◽  
Dong Ma ◽  
Zhe-Min Zhou
Keyword(s):  
Catalytic Activity ◽  
Nitrile Hydratase ◽  
Salt Bridge ◽  
Tunnel Entrance

The βGlu50 residue located adjacent to the substrate access tunnel entrance of nitrile hydratase from Pseudonocardia thermophila JCM3095 acts as an anchoring residue. Breaking the salt bridge between β50 residue...

Novel catalytic activity of nitrile hydratase from Rhodococcus sp. N771

2008 ◽  
Vol 106 (2) ◽  
pp. 174-179 ◽  
Author(s):  
Kayoko Taniguchi ◽  
Kensuke Murata ◽  
Yoshihiko Murakami ◽  
Shunya Takahashi ◽  
Takemichi Nakamura ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Nitrile Hydratase ◽  
Rhodococcus Sp

scholarly journals Characterization of iduronate sulphatase mutants affecting N-glycosylation sites and the cysteine-84 residue

1997 ◽  
Vol 326 (1) ◽  
pp. 243-247 ◽  
Author(s):  
Gilles MILLAT ◽  
Roseline FROISSART ◽  
Irène MAIRE ◽  
Dominique BOZON
Keyword(s):  
Catalytic Activity ◽  
Amino Acid Position ◽  
Cysteine Residue ◽  
Glycosylation Site ◽  
Site Directed Mutagenesis ◽  
Lysosomal Storage ◽  
Post Translational Modification ◽  
Cos Cells ◽  
Conserved Sequence ◽  
Glycosylation Sites

Iduronate sulphatase (IDS) is responsible for mucopolysaccharidosis type II, a rare recessive X-linked lysosomal storage disease. The aim of this work was to evaluate the functional importance of each N-glycosylation site, and of the cysteine-84 residue. IDS mutant cDNAs, lacking one of the eight potential N-glycosylation sites, were expressed in COS cells. Although each of the potential sites was used, none of the eight glycosylation sites appeared to be essential for lysosomal targeting. Another important sulphatase co- or post-translational modification for generating catalytic activity involves the conversion of a cysteine residue surrounded by a conserved sequence C-X-P-S-R into a 2-amino-3-oxopropionic acid residue [Schmidt, Selmer, Ingendoh and von Figura (1995) Cell 82, 271–278]. This conserved cysteine, located at amino acid position 84 in IDS, was replaced either by an alanine (C84A) or by a threonine (C84T) using site-directed mutagenesis. C84A and C84T mutant cDNAs were expressed either in COS cells or in human lymphoblastoid cells deleted for the IDS gene. C84A had a drastic effect both for IDS processing and for catalytic activity. The C84T mutation produced a small amount of mature forms but also abolished enzyme activity, confirming that the cysteine residue at position 84 is required for IDS activity.

scholarly journals Computational Design of Nitrile Hydratase from Pseudonocardia thermophila JCM3095 for Improved Thermostability

Molecules ◽  
2020 ◽  
Vol 25 (20) ◽  
pp. 4806
Author(s):  
Zhongyi Cheng ◽  
Yao Lan ◽  
Junling Guo ◽  
Dong Ma ◽  
Shijin Jiang ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Rational Design ◽  
Nitrile Hydratase ◽  
Elevated Temperatures ◽  
Residual Activity ◽  
Fold Increase ◽  
Computational Design ◽  
Site Directed Mutagenesis ◽  
Multiple Point ◽  
Substrate Affinity

High thermostability and catalytic activity are key properties for nitrile hydratase (NHase, EC 4.2.1.84) as a well-industrialized catalyst. In this study, rational design was applied to tailor the thermostability of NHase from Pseudonocardia thermophila JCM3095 (PtNHase) by combining FireProt server prediction and molecular dynamics (MD) simulation. Site-directed mutagenesis of non-catalytic residues provided by the rational design was subsequentially performed. The positive multiple-point mutant, namely, M10 (αI5P/αT18Y/αQ31L/αD92H/βA20P/βP38L/βF118W/βS130Y/βC189N/βC218V), was obtained and further analyzed. The Melting temperature (Tm) of the M10 mutant showed an increase by 3.2 °C and a substantial increase in residual activity of the enzyme at elevated temperatures was also observed. Moreover, the M10 mutant also showed a 2.1-fold increase in catalytic activity compared with the wild-type PtNHase. Molecular docking and MD simulations demonstrated better substrate affinity and improved thermostability for the mutant.

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