Bioluminescence emission of selected aequorin variants in an <i>in vivo</i> mouse model.

<p>Mice received a 5 μL intrastromal or antechamber injection in the right eye with variant aequorin and the left eye with HEPES Buffer. (A,B) Aequorin injected intrastromally at a concentration of 2.7 x 10⁻⁵ M, 30 s exposure. (C) Aequorin with L-4-methoxyphenylalanine at positions 82 and 86. (D) HEPES Buffer only in both eyes. (E) Aequorin injected in the antechamber at a concentration of 3.2 x 10⁻⁵ M, 60 s exposure. (F) Aequorin with L-4-iodophenylalanine at positions 82 and 86. Images generated with Matlab R2014b and LivingImage 4.4. Reprinted from [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0158579#pone.0158579.ref047" target="_blank">47</a>] under a CC BY license, with permission from <i>Open Access Dissertations</i>, original copyright 2015.</p

Emission characteristics of selected aequorin variants.

<p>Emission characteristics of selected aequorin variants.</p

Structures of wild-type aequorin and the non-natural amino acid analogs employed for site-selective incorporation into the protein.

<p>(A) Crystal structure of aequorin with the location of the Tyr82-His16-Trp86. The substituted tyrosine is at position 82 and the substituted tryptophan is at position 86. (B) Chemical structure of non-natural amino acids: L-4-aminophenylalanine, (C) L-4-bromophenylalanine, (D) L-4-iodophenylalanine, (E) L-4 methoxyphenylalanine (PDB ID: 1EJ3). Reprinted from [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0158579#pone.0158579.ref047" target="_blank">47</a>]under a CC BY license, with permission from <i>Open Access Dissertations</i>, original copyright 2015.</p

Structures of synthetic coelenterazines.

<p>Reprinted from [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0158579#pone.0158579.ref047" target="_blank">47</a>] under a CC BY license, with permission from <i>Open Access Dissertations</i>, original copyright 2015.</p

Emission wavelengths and bioluminescence half-lives of selected aequorin variants.

<p>(A) Bioluminescence emission spectra of aequorin with L-4-aminophenylalanine at position 86 with coelenterazine <i>cp</i> (red), aequorin with native coelenterazine (black), and L-4-methoxyphenylalanine at position 82 and 86 with coelenterazine <i>i</i> (green), illustrating the range of emission wavelengths in this study. (B) Half-life bioluminescence decay of aequorin with native coelenterazine (black), L-4-methoxyphenylalanine with coelenterazine <i>n</i> (orange), and L-4-iodophenylalanine with coelenterazine <i>i</i> (blue), illustrating the range of emission half-lives in this study. Reprinted from [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0158579#pone.0158579.ref047" target="_blank">47</a>] under a CC BY license, with permission from <i>Open Access Dissertations</i>, original copyright 2015.</p

Native coelenterazine in the binding pocket of aequorin.

<p>The grey dashed lines represent the Van der Waal surface of the ligand. Green dashed lines with an arrowhead represent an H-bond going from donor to recipient. The structure is based on aequorin’s crystal structure [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0158579#pone.0158579.ref043" target="_blank">43</a>](PDB ID: 1EJ3). Reprinted from [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0158579#pone.0158579.ref047" target="_blank">47</a>] under a CC BY license, with permission from <i>Open Access Dissertations</i>, original copyright 2015.</p