Mutagenesis study on the role of a lysine residue highly conserved in formate dehydrogenases and periplasmic nitrate reductases

Abstract The interaction between legumes and rhizobia leads to the establishment of a symbiotic relationship between plant and bacteria. This is characterized by the formation of a new organ, the nodule, which facilitates the fixation of atmospheric nitrogen (N2) by nitrogenase through the creation of a hypoxic environment. Nitric oxide (NO) accumulates at each stage of the symbiotic process. NO is involved in defense responses, nodule organogenesis and development, nitrogen fixation metabolism, and senescence induction. During symbiosis, either successively or simultaneously, NO regulates gene expression, modulates enzyme activities, and acts as a metabolic intermediate in energy regeneration processes via phytoglobin-NO respiration and the bacterial denitrification pathway. Due to the transition from normoxia to hypoxia during nodule formation, and the progressive presence of the bacterial partner in the growing nodules, NO production and degradation pathways change during the symbiotic process. This review analyzes the different source and degradation pathways of NO, and highlights the role of nitrate reductases and hemoproteins of both the plant and bacterial partners in the control of NO accumulation.

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A Mutagenesis Study to Investigate the Role of Alanine to Serine Mutations in the Adaptation of a Diatom Cytochrome c6 to Cold Temperatures

Biophysical Journal ◽

10.1016/j.bpj.2018.11.1837 ◽

2019 ◽

Vol 116 (3) ◽

pp. 337a

Author(s):

Miranda Wilson ◽

Jordyn Preusker ◽

Inaara Bhola ◽

Katherine Frato

Keyword(s):

Cytochrome C6 ◽

Cold Temperatures ◽

Mutagenesis Study

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Voltage-gated Na Channel Selectivity: The Role of the Conserved Domain III Lysine Residue

The Journal of General Physiology ◽

10.1085/jgp.200809991 ◽

2008 ◽

Vol 131 (6) ◽

pp. 523-529 ◽

Cited By ~ 35

Author(s):

Gregory M. Lipkind ◽

Harry A. Fozzard

Keyword(s):

Lysine Residue ◽

Na Channel ◽

Voltage Gated ◽

Conserved Domain ◽

Channel Selectivity ◽

Domain Iii

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Role of Acidic and Aromatic Amino Acids inRhodobacter CapsulatusCytochromec1. A Site-Directed Mutagenesis Study†

Biochemistry ◽

10.1021/bi020693r ◽

2003 ◽

Vol 42 (29) ◽

pp. 8818-8830 ◽

Cited By ~ 4

Author(s):

Jun Li ◽

Artur Osyczka ◽

Richard C. Conover ◽

Michael K. Johnson ◽

Hong Qin ◽

...

Keyword(s):

Amino Acids ◽

Aromatic Amino Acids ◽

Site Directed Mutagenesis ◽

Directed Mutagenesis ◽

A Site ◽

Mutagenesis Study

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Role of an invariant lysine residue in folate binding on Escherichia coli thymidylate synthase: Calorimetric and crystallographic analysis of the K48Q mutant

The International Journal of Biochemistry & Cell Biology ◽

10.1016/j.biocel.2008.02.025 ◽

2008 ◽

Vol 40 (10) ◽

pp. 2206-2217 ◽

Cited By ~ 7

Author(s):

Aldo A. Arvizu-Flores ◽

Rocio Sugich-Miranda ◽

Rodrigo Arreola ◽

Karina D. Garcia-Orozco ◽

Enrique F. Velazquez-Contreras ◽

...

Keyword(s):

Escherichia Coli ◽

Thymidylate Synthase ◽

Crystallographic Analysis ◽

Lysine Residue

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Abstract P117: A Conserved Trans-membrane Domain Lysine in the Integrin Beta1 Subunit Regulates Renal Collecting Duct Cell Function

Hypertension ◽

10.1161/hyp.68.suppl_1.p117 ◽

2016 ◽

Vol 68 (suppl_1) ◽

Author(s):

Sijo Mathew ◽

Jiang Chen ◽

Zhenwei Lu ◽

Charles Sanders ◽

Roy Zent

Keyword(s):

Cell Function ◽

Collecting Duct ◽

Duct Cell ◽

Lysine Residue ◽

Mutant Protein ◽

Integrin Β1 ◽

Binding Ability ◽

Integrin Β3 ◽

Collecting Duct Cells

Integrins are heterodimeric trans-membrane receptor proteins that mediate the interaction of cells with extracellular matrix proteins (ECM). They mediate various growth factor dependent cell-signaling pathways during the development of fibrosis that is characteristic of all forms of chronic kidney diseases. Although integrin β1 is the most abundant integrin subunit in kidney and can form complexes with 12 different α subunits, integrin β3 is the best studied integrin β subunit and serves as the canonical model for integrin function based on the high sequence homology between the trans-membrane and cytosolic domains of integrin β subunits. A conserved lysine residue towards the C-terminus of integrin β3 subunit is reported to be important for regulating the activation of integrin αIIbβ3 complexes; however the functional importance of this lysine is unknown in β1 integrins. We investigated the role of this lysine residue in integrin β1-dependent kidney collecting duct cell function. We expressed the mutant protein where the lysine is mutated to glutamic acid in collecting duct cells null for integrin β1. Collecting duct cells expressing mutant protein had decreased the adhesion of cells to collagen IV mediated by integrin α1β1 by 80% (0.95 vs 0.18). This mutation also decreased the ability of IMCD cells adhesion to collagen I mediated by integrin α2β1 by 82% (0.78 vs 0.15). In contrast to earlier reports in integrin β3, this mutation did not significantly alter the amount of active integrin β1 on the cell surface as estimated by FACS analysis; however we did observe a decrease in conformation specific antibody binding on cells adhered to collagen (0.70 vs 0.30). We also investigated the role of this lysine residue in complex formation of purified integrin β1 with integrin α1 and α2 TM/CT domains in phospholipid bicelles using fluorescence anisotropy. The dissociation constant for binding was estimated to be >3.2 mol and mutation of lysine residue did not significantly alter their binding ability. This contrasted with integrin αIIb β3 where we found fourfold decrease in binding ability (Kd 0.09 ± 0.03 mol and 0.33 ± 0.05 mol). Our data clearly suggest that conserved transmembrane lysine in both integrin β3 and integrin β1 regulate cell functions by distinct mechanisms.

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Role of a Helix B Lysine Residue in the Photoactive Site in Channelrhodopsins

Biophysical Journal ◽

10.1016/j.bpj.2014.03.002 ◽

2014 ◽

Vol 106 (8) ◽

pp. 1607-1617 ◽

Cited By ~ 12

Author(s):

Hai Li ◽

Elena G. Govorunova ◽

Oleg A. Sineshchekov ◽

John L. Spudich

Keyword(s):

Lysine Residue

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Role of positive charge of lysine residue on cytochrome c3 for electrostatic interaction with hydrogenase

Journal of Porphyrins and Phthalocyanines ◽

10.1142/s1088424607000096 ◽

2007 ◽

Vol 11 (01) ◽

pp. 66-73

Author(s):

Shin Iida ◽

Noriyuki Asakura ◽

Kenji Tabata ◽

Ichiro Okura ◽

Toshiaki Kamachi

Keyword(s):

Electron Transfer ◽

Cytochrome C ◽

Hydrogen Evolution ◽

Electrostatic Interaction ◽

Positive Charge ◽

Lysine Residue ◽

Site Directed Mutagenesis ◽

Desulfovibrio Vulgaris ◽

Lysine Residues

Cytochrome c3 from Desulfovibrio vulgaris (Miyazaki) is an electron transfer protein containing four hemes per molecule. Its physiological electron transfer partner is the hydrogenase which catalyzes reversible oxidation of hydrogen. The complex formation between cytochrome c3 and hydrogenase is caused by electrostatic interaction, because cytochrome c3 is a basic protein and hydrogenase is an acidic protein. As cytochrome c3 has 20 lysine residues among 108 amino acids, the positive charges of some lysine residues may play an important role in the interaction with hydrogenase. To clarify the role of positive charge of lysine residue, the positive charge was changed to neutral or negative charge using chemical modification and site-directed mutagenesis. When the positive charges around heme IV were changed, the hydrogen evolution rate with hydrogenase decreased. The affinity between hydrogenase and mutated cytochrome c3 (K57Q, K57E, K72Q, K94Q, K94E) were not affected. On the other hand, the affinity of K72E cytochrome c3 for hydrogenase was very low. These results suggest that the positive charge around heme IV plays an important role in the electrostatic interaction with hydrogenase in hydrogen evolution.

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