scholarly journals Noncanonical Amino Acid Labeling in Vivo to Visualize and Affinity Purify Newly Synthesized Proteins in Larval Zebrafish

2011 ◽  
Vol 3 (1) ◽  
pp. 40-49 ◽  
Author(s):  
Flora I. Hinz ◽  
Daniela C. Dieterich ◽  
David A. Tirrell ◽  
Erin M. Schuman
Keyword(s):  
Amino Acid ◽  
Larval Zebrafish ◽  
Amino Acid Labeling ◽  
Noncanonical Amino Acid

scholarly journals BIOSYNTHESIS IN ISOLATED ACETABULARIA CHLOROPLASTS

1972 ◽  
Vol 54 (2) ◽  
pp. 279-294 ◽  
Author(s):  
David C. Shephard ◽  
Wendy B. Levin
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Amino Acid ◽  
Technical Problems ◽  
Hydrolyzed Protein ◽  
Protein Amino Acids ◽  
Amino Acid Labeling ◽  
Labeling Pattern

The ability of chloroplasts isolated from Acetabulana mediterranea to synthesize the protein amino acids has been investigated. When this chloroplast isolate was presented with 14CO2 for periods of 6–8 hr, tracer was found in essentially all amino acid species of their hydrolyzed protein Phenylalanine labeling was not detected, probably due to technical problems, and hydroxyproline labeling was not tested for The incorporation of 14CO2 into the amino acids is driven by light and, as indicated by the amount of radioactivity lost during ninhydrin decarboxylation on the chromatograms, the amino acids appear to be uniformly labeled. The amino acid labeling pattern of the isolate is similar to that found in plastids labeled with 14CO2 in vivo. The chloroplast isolate did not utilize detectable amounts of externally supplied amino acids in light or, with added adenosine triphosphate (ATP), in darkness. It is concluded that these chloroplasts are a tight cytoplasmic compartment that is independent in supplying the amino acids used for its own protein synthesis. These results are discussed in terms of the role of contaminants in the observed synthesis, the "normalcy" of Acetabularia chloroplasts, the synthetic pathways for amino acids in plastids, and the implications of these observations for cell compartmentation and chloroplast autonomy.

scholarly journals Accurate Determination of Human CPR Conformational Equilibrium by smFRET Using Dual Orthogonal Noncanonical Amino Acid Labeling

ChemBioChem ◽  
2019 ◽  
Vol 20 (5) ◽  
pp. 659-666 ◽  
Author(s):  
Robert B. Quast ◽  
Fataneh Fatemi ◽  
Michel Kranendonk ◽  
Emmanuel Margeat ◽  
Gilles Truan
Keyword(s):  
Amino Acid ◽  
Conformational Equilibrium ◽  
Accurate Determination ◽  
Amino Acid Labeling ◽  
Noncanonical Amino Acid

In Vivo Termini Amino Acid Labeling for Quantitative Proteomics

2011 ◽  
Vol 83 (15) ◽  
pp. 6026-6033 ◽  
Author(s):  
Ai-Ying Nie ◽  
Lei Zhang ◽  
Guo-Quan Yan ◽  
Jun Yao ◽  
Yang Zhang ◽  
...  
Keyword(s):  
Amino Acid ◽  
Quantitative Proteomics ◽  
Amino Acid Labeling

Global in vivo terminal amino acid labeling for exploring differential expressed proteins induced by dialyzed serum cultivation

The Analyst ◽  
2014 ◽  
Vol 139 (18) ◽  
pp. 4497-4504 ◽  
Author(s):  
Li-Qi Xie ◽  
Ai-Ying Nie ◽  
Shu-Jun Yang ◽  
Chao Zhao ◽  
Lei Zhang ◽  
...  
Keyword(s):  
Amino Acid ◽  
High Throughput ◽  
Trypsin Digestion ◽  
Metabolic Labeling ◽  
Terminal Amino Acid ◽  
Terminal Amino ◽  
Amino Acid Labeling ◽  
Dialyzed Serum

An accurate and high throughput isobaric MS2 quantification strategy based on metabolic labeling and trypsin digestion.

scholarly journals Mechanism and site of action of big dynorphin on ASIC1a

2020 ◽  
Vol 117 (13) ◽  
pp. 7447-7454 ◽  
Author(s):  
Christian B. Borg ◽  
Nina Braun ◽  
Stephanie A. Heusser ◽  
Yasmin Bay ◽  
Daniel Weis ◽  
...  
Keyword(s):  
Amino Acids ◽  
Ischemic Stroke ◽  
Amino Acid ◽  
Binding Site ◽  
Neuronal Death ◽  
Electrostatic Interactions ◽  
Rational Design ◽  
Site Of Action ◽  
Noncanonical Amino Acid

Acid-sensing ion channels (ASICs) are proton-gated cation channels that contribute to neurotransmission, as well as initiation of pain and neuronal death following ischemic stroke. As such, there is a great interest in understanding the in vivo regulation of ASICs, especially by endogenous neuropeptides that potently modulate ASICs. The most potent endogenous ASIC modulator known to date is the opioid neuropeptide big dynorphin (BigDyn). BigDyn is up-regulated in chronic pain and increases ASIC-mediated neuronal death during acidosis. Understanding the mechanism and site of action of BigDyn on ASICs could thus enable the rational design of compounds potentially useful in the treatment of pain and ischemic stroke. To this end, we employ a combination of electrophysiology, voltage-clamp fluorometry, synthetic BigDyn analogs, and noncanonical amino acid-mediated photocrosslinking. We demonstrate that BigDyn binding results in an ASIC1a closed resting conformation that is distinct from open and desensitized states induced by protons. Using alanine-substituted BigDyn analogs, we find that the BigDyn modulation of ASIC1a is primarily mediated through electrostatic interactions of basic amino acids in the BigDyn N terminus. Furthermore, neutralizing acidic amino acids in the ASIC1a extracellular domain reduces BigDyn effects, suggesting a binding site at the acidic pocket. This is confirmed by photocrosslinking using the noncanonical amino acid azidophenylalanine. Overall, our data define the mechanism of how BigDyn modulates ASIC1a, identify the acidic pocket as the binding site for BigDyn, and thus highlight this cavity as an important site for the development of ASIC-targeting therapeutics.

Site Specific Incorporation of Amino Acid Analogues into Proteins In Vivo

2000 ◽  
Author(s):  
Anne K. Kowal ◽  
Caroline Kohrer ◽  
Uttam L. RajBhandary
Keyword(s):  
Amino Acid ◽  
Site Specific ◽  
Amino Acid Analogues

Oxytocin analogues with non-coded amino acid residues in position 8: [8-Neopentylglycine]oxytocin and [8-cycloleucine]oxytocin

1987 ◽  
Vol 52 (9) ◽  
pp. 2317-2325 ◽  
Author(s):  
Jan Hlaváček ◽  
Jan Pospíšek ◽  
Jiřina Slaninová ◽  
Walter Y. Chan ◽  
Victor J. Hruby
Keyword(s):  
Amino Acid ◽  
Solid Phase Synthesis ◽  
Solid Phase ◽  
The Other ◽  
Amino Acid Residues ◽  
Phase Synthesis ◽  
Other Hand ◽  
Fragment Condensation

[8-Neopentylglycine]oxytocin (II) and [8-cycloleucine]oxytocin (III) were prepared by a combination of solid-phase synthesis and fragment condensation. Both analogues exhibited decreased uterotonic potency in vitro, each being about 15-30% that of oxytocin. Analogue II also displayed similarly decreased uterotonic potency in vivo and galactogogic potency. On the other hand, analogue III exhibited almost the same potency as oxytocin in the uterotonic assay in vivo and in the galactogogic assay.

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