Measurement of Nucleotide Hydrolysis Using Fluorescent Biosensors for Phosphate

The second messenger 3′,5′-cyclic nucleoside adenosine monophosphate (cAMP) plays a key role in signal transduction across prokaryotes and eukaryotes. Cyclic AMP signaling is compartmentalized into microdomains to fulfil specific functions. To define the function of cAMP within these microdomains, signaling needs to be analyzed with spatio-temporal precision. To this end, optogenetic approaches and genetically encoded fluorescent biosensors are particularly well suited. Synthesis and hydrolysis of cAMP can be directly manipulated by photoactivated adenylyl cyclases (PACs) and light-regulated phosphodiesterases (PDEs), respectively. In addition, many biosensors have been designed to spatially and temporarily resolve cAMP dynamics in the cell. This review provides an overview about optogenetic tools and biosensors to shed light on the subcellular organization of cAMP signaling.

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Distinct kinetics for nucleotide hydrolysis in lymphocytes isolated from blood, spleen and cervical lymph nodes: Characterization of ectonucleotidase activity

Cell Biochemistry and Function ◽

10.1002/cbf.3616 ◽

2021 ◽

Author(s):

Pedro H. Doleski ◽

Fernanda L. Cabral ◽

Stephen A. Adefegha ◽

Matheus H. Jantsch ◽

Renan S. Ebone ◽

...

Keyword(s):

Lymph Nodes ◽

Cervical Lymph Nodes ◽

Nucleotide Hydrolysis

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Simulation of extracellular nucleotide hydrolysis and determination of kinetic constants for the ectonucleotidases.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)66741-9 ◽

1986 ◽

Vol 261 (33) ◽

pp. 15505-15507

Author(s):

L L Slakey ◽

K Cosimini ◽

J P Earls ◽

C Thomas ◽

E L Gordon

Keyword(s):

Kinetic Constants ◽

Extracellular Nucleotide ◽

Nucleotide Hydrolysis

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New Fluorescent Biosensors for Amiloid Fibriles Detection

2018 IEEE 8th International Conference Nanomaterials: Application & Properties (NAP) ◽

10.1109/nap.2018.8914772 ◽

2018 ◽

Author(s):

A.A. Lugovski ◽

M.P. Samtsov ◽

A.P. Lugovski ◽

E.S. Voropay ◽

A.V. Lavysh ◽

...

Keyword(s):

Fluorescent Biosensors

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Aqueous extract of Ilex paraguariensis decreases nucleotide hydrolysis in rat blood serum

Journal of Ethnopharmacology ◽

10.1016/j.jep.2004.10.015 ◽

2005 ◽

Vol 97 (1) ◽

pp. 73-77 ◽

Cited By ~ 17

Author(s):

Milena Görgen ◽

Kátia Turatti ◽

Afonso R. Medeiros ◽

Andréia Buffon ◽

Carla D. Bonan ◽

...

Keyword(s):

Blood Serum ◽

Aqueous Extract ◽

Ilex Paraguariensis ◽

Nucleotide Hydrolysis ◽

Rat Blood

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Molecular motors--a paradigm for mutant analysis

Journal of Cell Science ◽

10.1242/jcs.113.8.1311 ◽

2000 ◽

Vol 113 (8) ◽

pp. 1311-1318 ◽

Cited By ~ 1

Author(s):

S.A. Endow

Keyword(s):

Molecular Motors ◽

Protein Function ◽

Rational Design ◽

Atp Hydrolysis ◽

Unique Property ◽

Genetic Screens ◽

Nucleotide Hydrolysis ◽

Mutant Analysis ◽

Motor Activities ◽

Natural Variants

Molecular motors perform essential functions in the cell and have the potential to provide insights into the basis of many important processes. A unique property of molecular motors is their ability to convert energy from ATP hydrolysis into work, enabling the motors to bind to and move along cytoskeletal filaments. The mechanism of energy conversion by molecular motors is not yet understood and may lead to the discovery of new biophysical principles. Mutant analysis could provide valuable information, but it is not obvious how to obtain mutants that are informative for study. The analysis presented here points out several strategies for obtaining mutants by selection from molecular or genetic screens, or by rational design. Mutants that are expected to provide important information about the motor mechanism include ATPase mutants, which interfere with the nucleotide hydrolysis cycle, and uncoupling mutants, which unlink basic motor activities and reveal their interdependence. Natural variants can also be exploited to provide unexpected information about motor function. This general approach to uncovering protein function by analysis of informative mutants is applicable not only to molecular motors, but to other proteins of interest.

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A key tyrosine substitution restricts nucleotide hydrolysis by the ectoenzyme NPP5

FEBS Journal ◽

10.1111/febs.14266 ◽

2017 ◽

Cited By ~ 3

Author(s):

Alexei Gorelik ◽

Antsa Randriamihaja ◽

Katalin Illes ◽

Bhushan Nagar

Keyword(s):

Nucleotide Hydrolysis

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Modulation of intestinal sulfur assimilation metabolism regulates iron homeostasis

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1715302115 ◽

2018 ◽

Vol 115 (12) ◽

pp. 3000-3005 ◽

Cited By ~ 7

Author(s):

Benjamin H. Hudson ◽

Andrew T. Hale ◽

Ryan P. Irving ◽

Shenglan Li ◽

John D. York

Keyword(s):

Iron Deficiency ◽

Iron Deficiency Anemia ◽

Iron Reduction ◽

Genetic Basis ◽

Iron Homeostasis ◽

Deficiency Anemia ◽

Sulfur Assimilation ◽

Nucleotide Hydrolysis ◽

Evolutionarily Conserved ◽

Organismal Development

Sulfur assimilation is an evolutionarily conserved pathway that plays an essential role in cellular and metabolic processes, including sulfation, amino acid biosynthesis, and organismal development. We report that loss of a key enzymatic component of the pathway, bisphosphate 3′-nucleotidase (Bpnt1), in mice, both whole animal and intestine-specific, leads to iron-deficiency anemia. Analysis of mutant enterocytes demonstrates that modulation of their substrate 3′-phosphoadenosine 5′-phosphate (PAP) influences levels of key iron homeostasis factors involved in dietary iron reduction, import and transport, that in part mimic those reported for the loss of hypoxic-induced transcription factor, HIF-2α. Our studies define a genetic basis for iron-deficiency anemia, a molecular approach for rescuing loss of nucleotidase function, and an unanticipated link between nucleotide hydrolysis in the sulfur assimilation pathway and iron homeostasis.

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