Dimerization of the plant molybdenum insertase Cnx1E is required for synthesis of the molybdenum cofactor

2016 ◽  
Vol 474 (1) ◽  
pp. 163-178 ◽  
Author(s):  
Joern Krausze ◽  
Corinna Probst ◽  
Ute Curth ◽  
Joachim Reichelt ◽  
Sayantan Saha ◽  
...  
Keyword(s):  
Amino Acid ◽  
Biochemical Characterization ◽  
Molybdenum Cofactor ◽  
Transfer Reactions ◽  
Single Amino Acid ◽  
Amino Acid Exchange ◽  
Prosthetic Group ◽  
Redox Active ◽  
Acid Exchange

The molybdenum cofactor (Moco) is a redox active prosthetic group, essentially required for numerous enzyme-catalyzed two electron transfer reactions. Moco is synthesized by an evolutionarily old and highly conserved multistep pathway. In the last step of Moco biosynthesis, the molybdenum center is inserted into the final Moco precursor adenylated molybdopterin (MPT-AMP). This unique and yet poorly characterized maturation reaction finally yields physiologically active Moco. In the model plant Arabidopsis, the two domain enzyme, Cnx1, is required for Moco formation. Recently, a genetic screen identified novel Arabidopsis cnx1 mutant plant lines each harboring a single amino acid exchange in the N-terminal Cnx1E domain. Biochemical characterization of the respective recombinant Cnx1E variants revealed two different amino acid exchanges (S197F and G175D) that impair Cnx1E dimerization, thus linking Cnx1E oligomerization to Cnx1 functionality. Analysis of the Cnx1E structure identified Cnx1E active site-bound molybdate and magnesium ions, which allowed to fine-map the Cnx1E MPT-AMP-binding site.

scholarly journals Collapse of the native structure caused by a single amino acid exchange in human NAD (P)H:quinone oxidoreductase 1

FEBS Journal ◽  
2014 ◽  
Vol 281 (20) ◽  
pp. 4691-4704 ◽  
Author(s):  
Wolf‐Dieter Lienhart ◽  
Venugopal Gudipati ◽  
Michael K. Uhl ◽  
Alexandra Binter ◽  
Sergio A. Pulido ◽  
...  
Keyword(s):  
Amino Acid ◽  
Single Amino Acid ◽  
Native Structure ◽  
Amino Acid Exchange ◽  
Acid Exchange

scholarly journals A Single Amino Acid Exchange Inverts Susceptibility of Related Receptor Tyrosine Kinases for the ATP Site Inhibitor STI-571

2002 ◽  
Vol 278 (7) ◽  
pp. 5148-5155 ◽  
Author(s):  
Frank D. Böhmer ◽  
Luchezar Karagyozov ◽  
Andrea Uecker ◽  
Hubert Serve ◽  
Alexander Botzki ◽  
...  
Keyword(s):  
Amino Acid ◽  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Single Amino Acid ◽  
Amino Acid Exchange ◽  
Sti 571 ◽  
Acid Exchange

A single amino acid exchange abolishes dimerization of the androgen receptor and causes Reifenstein syndrome

1995 ◽  
Vol 111 (1) ◽  
pp. 93-98 ◽  
Author(s):  
Andreas Gast ◽  
Felizia Neuschmid-Kaspar ◽  
Helmut Klocker ◽  
Andrew C.B. Cato
Keyword(s):  
Androgen Receptor ◽  
Amino Acid ◽  
Single Amino Acid ◽  
Amino Acid Exchange ◽  
Acid Exchange

scholarly journals Exploring the full natural diversity of single amino acid exchange reveals that 40–60% of BSLA positions improve organic solvents resistance

2018 ◽  
Vol 5 (1) ◽  
Author(s):  
Victorine Josiane Frauenkron-Machedjou ◽  
Alexander Fulton ◽  
Jing Zhao ◽  
Lina Weber ◽  
Karl-Erich Jaeger ◽  
...  
Keyword(s):  
Amino Acid ◽  
Organic Solvents ◽  
Single Amino Acid ◽  
Amino Acid Exchange ◽  
Natural Diversity ◽  
Acid Exchange

scholarly journals Characterization of fibrinogen Milano I: amino acid exchange gamma 330 Asp----Val impairs fibrin polymerization

Blood ◽  
1986 ◽  
Vol 67 (6) ◽  
pp. 1751-1756
Author(s):  
P Reber ◽  
M Furlan ◽  
C Rupp ◽  
M Kehl ◽  
A Henschen ◽  
...  
Keyword(s):  
Amino Acid ◽  
Time Course ◽  
Precipitation Method ◽  
Functional Assay ◽  
Amino Acid Exchange ◽  
Gamma Chain ◽  
Tentative Diagnosis ◽  
Fibrin Monomers ◽  
Acid Exchange

An abnormal fibrinogen was found in two asymptomatic members (father and daughter) of the same family, originating from northern Italy. Routine coagulation studies revealed prolonged thrombin and reptilase clotting times. Plasma fibrinogen levels, as determined by a functional assay, were markedly diminished, whereas the heat precipitation method indicated normal fibrinogen values. On the basis of these findings, a tentative diagnosis of dysfibrinogenemia was made, and according to the accepted nomenclature, this fibrinogen variant was called “fibrinogen Milano l.” The time course of fibrinopeptide A and B release from fibrinogen Milano l was normal, but the aggregation of fibrin monomers was delayed. Two-dimensional electrophoresis of reduced variant fibrinogen chains showed a defective gamma-chain with increased cathodic mobility. An abnormal electrophoretic mobility was observed also for the gamma-chain remnants of fibrinogen fragments D1 and D2 derived from fibrinogen Milano l, whereas the charge anomaly was lost after a further digestion by plasmin to D3, suggesting that the structure abnormality of this variant is situated in the region gamma 303–356. An abnormal peptide was isolated after cyanogen bromide cleavage of intact fibrinogen Milano l. This fragment spans from position gamma 311 to gamma 336. Amino acid analysis of the abnormal peptide showed the presence of valine and a diminished content of aspartic acid. Sequence analysis demonstrated an amino acid exchange Asp----Val in the gamma-chain at position 330.

scholarly journals A Single Amino Acid Exchange, Arg-45 to Ala, Generates an Epidermal Growth Factor (EGF) Mutant with High Affinity for the Chicken EGF Receptor

1995 ◽  
Vol 270 (38) ◽  
pp. 22337-22343 ◽  
Author(s):  
Monique L. M. van de Poll ◽  
Anne E. G. Lenferink ◽  
Marianne J. H. van Vugt ◽  
Jacqueline J. L. Jacobs ◽  
Jannie W. H. Janssen ◽  
...  
Keyword(s):  
Amino Acid ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Egf Receptor ◽  
Single Amino Acid ◽  
Amino Acid Exchange ◽  
High Affinity ◽  
Epidermal Growth ◽  
Acid Exchange

scholarly journals Biochemical Characterization of Molybdenum Cofactor-free Nitrate Reductase from Neurospora crassa

2013 ◽  
Vol 288 (20) ◽  
pp. 14657-14671 ◽  
Author(s):  
Phillip Ringel ◽  
Joern Krausze ◽  
Joop van den Heuvel ◽  
Ute Curth ◽  
Antonio J. Pierik ◽  
...  
Keyword(s):  
Nitrate Reductase ◽  
Neurospora Crassa ◽  
Biochemical Characterization ◽  
Nitrate Assimilation ◽  
Molybdenum Cofactor ◽  
Site Formation ◽  
Epr Spectrum ◽  
Redox Active

Nitrate reductase (NR) is a complex molybdenum cofactor (Moco)-dependent homodimeric metalloenzyme that is vitally important for autotrophic organism as it catalyzes the first and rate-limiting step of nitrate assimilation. Beside Moco, eukaryotic NR also binds FAD and heme as additional redox active cofactors, and these are involved in electron transfer from NAD(P)H to the enzyme molybdenum center where reduction of nitrate to nitrite takes place. We report the first biochemical characterization of a Moco-free eukaryotic NR from the fungus Neurospora crassa, documenting that Moco is necessary and sufficient to induce dimer formation. The molybdenum center of NR reconstituted in vitro from apo-NR and Moco showed an EPR spectrum identical to holo-NR. Analysis of mutants unable to bind heme or FAD revealed that insertion of Moco into NR occurs independent from the insertion of any other NR redox cofactor. Furthermore, we showed that at least in vitro the active site formation of NR is an autonomous process.

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