WO2014016455A1 - Variants de la protéine de fusion sensible au calcium tdtomato-aequorine - Google Patents

Variants de la protéine de fusion sensible au calcium tdtomato-aequorine Download PDF

Info

Publication number
WO2014016455A1
WO2014016455A1 PCT/ES2013/070484 ES2013070484W WO2014016455A1 WO 2014016455 A1 WO2014016455 A1 WO 2014016455A1 ES 2013070484 W ES2013070484 W ES 2013070484W WO 2014016455 A1 WO2014016455 A1 WO 2014016455A1
Authority
WO
WIPO (PCT)
Prior art keywords
protein
seq
cells
cell
subunit
Prior art date
Application number
PCT/ES2013/070484
Other languages
English (en)
Spanish (es)
Inventor
Adil BAKAYAN
Juan Francisco LLOPIS BORRÁS
Original Assignee
Universidad De Castilla La Mancha
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Universidad De Castilla La Mancha filed Critical Universidad De Castilla La Mancha
Publication of WO2014016455A1 publication Critical patent/WO2014016455A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y113/00—Oxidoreductases acting on single donors with incorporation of molecular oxygen (oxygenases) (1.13)
    • C12Y113/12—Oxidoreductases acting on single donors with incorporation of molecular oxygen (oxygenases) (1.13) with incorporation of one atom of oxygen (internal monooxygenases or internal mixed function oxidases)(1.13.12)
    • C12Y113/12005—Renilla-luciferin 2-monooxygenase (1.13.12.5), i.e. renilla-luciferase
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
    • C07K14/43595—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from coelenteratae, e.g. medusae
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0069—Oxidoreductases (1.) acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6872—Intracellular protein regulatory factors and their receptors, e.g. including ion channels
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/60—Fusion polypeptide containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/61—Fusion polypeptide containing an enzyme fusion for detection (lacZ, luciferase)

Abstract

La présente invention se rapporte à des protéines dérivées de la protéine de fusion tdTomato-Aequorine, à une composition qui les contient, à la séquence nucléotidique qui les code, à son utilisation pour la détection et/ou la quantification in vitro de la concentration de calcium intracellulaire, ainsi qu'à un kit qui les contient et à l'utilisation dudit kit à cette même fin. L'invention concerne également le procédé d'obtention de la protéine de fusion de l'invention. Elle porte en outre sur un procédé de criblage comprenant l'utilisation des protéines de fusion de la présente invention.
PCT/ES2013/070484 2012-07-27 2013-07-08 Variants de la protéine de fusion sensible au calcium tdtomato-aequorine WO2014016455A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ES201231220A ES2445018B1 (es) 2012-07-27 2012-07-27 VARIANTES DE LA PROTEÍNA DE FUSIÓN SENSIBLE AL CALCIO tdTOMATO-AEQUORINA
ESP201231220 2012-07-27

Publications (1)

Publication Number Publication Date
WO2014016455A1 true WO2014016455A1 (fr) 2014-01-30

Family

ID=49996645

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/ES2013/070484 WO2014016455A1 (fr) 2012-07-27 2013-07-08 Variants de la protéine de fusion sensible au calcium tdtomato-aequorine

Country Status (2)

Country Link
ES (1) ES2445018B1 (fr)
WO (1) WO2014016455A1 (fr)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001092300A2 (fr) * 2000-06-01 2001-12-06 Institut Pasteur Aequorine-gfp chimere utilisee comme reporter de ca++ bioluminescent au niveau de la cellule unitaire
US20100233726A1 (en) * 1996-01-31 2010-09-16 The Regents Of The University Of California Tandem fluorescent protein constructs

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100233726A1 (en) * 1996-01-31 2010-09-16 The Regents Of The University Of California Tandem fluorescent protein constructs
WO2001092300A2 (fr) * 2000-06-01 2001-12-06 Institut Pasteur Aequorine-gfp chimere utilisee comme reporter de ca++ bioluminescent au niveau de la cellule unitaire

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
BAKAYAN, A. ET AL.: "Red fluorescent protein-aequorin fusions as improved bioluminescent Ca2+ reporters in single cells and mice", PLOS ONE, vol. 6, no. 5, 2011, pages E19520 *
CURIE, T. ET AL.: "Red-shifted aequorin-based bioluminescent reporters for in vivo imagingof Ca2 signaling", MOLECULAR IMAGING, vol. 6, no. 1, 2007, pages 30 - 42 *
DE, A. ET AL.: "BRET3: a red-shifted bioluminescence resonance energy transfer (BRET)-based integrated platform for imaging protein-protein interactions from single live cells and living animals", FASEB JOURNAL, vol. 23, no. 8, 2009, pages 2702 - 2709 *
DIKICI, E. ET AL.: "Aequorin variants with improved bioluminescence properties", PROTEIN ENGINEERING DESIGN & SELECTION, vol. 22, no. 4, 2009, pages 243 - 248 *
EVERS, T.H. ET AL.: "Quantitative understanding of the energy transfer between fluorescent proteins connected via flexible peptide linkers", BIOCHEMISTRY, vol. 45, no. 44, 2006, pages 13183 - 13192 *
SHIMOZONO, S. ET AL.: "Concatenation of cyan and yellow fluorescent proteins for efficient resonance energy transfer", BIOCHEMISTRY, vol. 45, no. 20, 2006, pages 6267 - 6271 *
TRICOIRE, L. ET AL.: "Calcium dependence of aequorin bioluminescence dissected by random mutagenesis", PROCEEDINGS OF THE NATIONAL ACADEMYO SCIENCES, vol. 103, no. 25, 2006, U S A, pages 9500 - 9505 *
TSUZUKI, K. ET AL.: "Thermostable mutants of the photoprotein aequorin obtained by in vitroevolution", JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 280, no. 40, 2005, pages 34324 - 34331 *
WEBB, S.E. ET AL.: "The use of aequorins to record and visualize Ca(2+) dynamics: from subcellular microdomains to whole organisms", METHODS IN CELL BIOLOGY, vol. 99, 2010, pages 263 - 300 *

Also Published As

Publication number Publication date
ES2445018A1 (es) 2014-02-27
ES2445018B1 (es) 2014-12-12

Similar Documents

Publication Publication Date Title
US10800829B2 (en) Optogenetic probes for measuring membrane potential
CA3157995A1 (fr) Composes indicateurs, dispositifs comprenant des composes indicateurs, et leurs procedes de fabrication et d'utilisation
ES2237643T3 (es) Fotoproteina con bioluminiscencia mejorada.
US9908918B2 (en) Cyan-excitable orange-red fluorescent proteins and bioluminescent resonance energy transfer systems
ES2399563T3 (es) Proteínas verdes fluorescentes modificadas y método para la utilización de las mismas
EP3080165A1 (fr) Récepteurs optiquement activés
JPWO2010119721A1 (ja) 超高輝度で安定な人工生物発光酵素
JP2010046084A (ja) Ａｅｑｕｏｒｅａｃｏｅｒｕｌｅｓｃｅｎｓ由来の新規な蛍光タンパク質およびその使用方法
US20240027344A1 (en) Fluorescent Probe for Branched Chain Amino Acids and Use Thereof
US20150225467A1 (en) Novel far red fluorescent protein
US8524447B2 (en) Fluorescently labeled fusion protein for assaying adenosine triphosphate
EP3037534A1 (fr) Polypeptide présentant des propriétés fluorescentes et son utilisation
ES2445018B1 (es) VARIANTES DE LA PROTEÍNA DE FUSIÓN SENSIBLE AL CALCIO tdTOMATO-AEQUORINA
EP2436690B1 (fr) Polypeptides réversiblement photocommutables
ES2329666T3 (es) Proteinas fluorescentes de maduracion rapida y procedimientos de uso de las mismas.
KR20130103560A (ko) 개변 형광 단백질
WO2017057752A1 (fr) Nouvelles enzymes bioluminescentes artificielles
WO2013163681A1 (fr) Protéines fluorescentes et leurs utilisations
WO2016038750A1 (fr) Luciférase recombinée de type fractionné et procédé d'analyse l'utilisant
ES2452485A1 (es) Mutantes de apoacuorina y metodos para su uso
Kim Molecular Engineering of Monomeric Red Fluorescent Proteins for Biosensing Applications
RU2338785C2 (ru) Флуоресцирующие белки и хромопротеины из видов hydrozoa, не относящихся к aequorea, и способы их получения
Demirbaş Investigating the homo-dimerization of ADGRD1/GPR133 and ADGRG5/GPR114 via resonance energy transfer techniques
ES2427854B1 (es) Sensores de calcio y métodos para la detección de calcio libre intracelular
JP2012085542A (ja) 特定部位のアミノ酸を置換した緑色蛍光蛋白質又はそのホモログを用いたカルシウムセンサー蛋白質

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 13823567

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 13823567

Country of ref document: EP

Kind code of ref document: A1