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Tytuł:
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Green fluorescent protein-based lactate and pyruvate indicators suitable for biochemical assays and live cell imaging.
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Autorzy:
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Harada K; Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo, 153-8902, Japan.
Chihara T; Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo, 113-0033, Japan.
Hayasaka Y; Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo, 153-8902, Japan.
Mita M; Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo, 153-8902, Japan.
Takizawa M; Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo, 153-8902, Japan.
Ishida K; Myoridge Co. Ltd., 46-29 Yoshidashimoadachi-cho, Sakyo, Kyoto, 606-8501, Japan.
Arai M; Department of Biology, Faculty of Science, Kyushu University, 774 Motooka, Nishi, Fukuoka, 819-0395, Japan.
Tsuno S; Dairy Science and Technology Institute, Kyodo Milk Industry Co. Ltd., Hinode-machi, Nishitama-gun, Tokyo, 190-0182, Japan.
Matsumoto M; Dairy Science and Technology Institute, Kyodo Milk Industry Co. Ltd., Hinode-machi, Nishitama-gun, Tokyo, 190-0182, Japan.
Ishihara T; Department of Biology, Faculty of Science, Kyushu University, 774 Motooka, Nishi, Fukuoka, 819-0395, Japan.
Ueda H; Laboratory for Chemistry and Life Science, Institute of Innovative Research, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama, Kanagawa, 226-8503, Japan.
Kitaguchi T; Laboratory for Chemistry and Life Science, Institute of Innovative Research, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama, Kanagawa, 226-8503, Japan. .
Tsuboi T; Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo, 153-8902, Japan. .; Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo, 113-0033, Japan. .
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Źródło:
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Scientific reports [Sci Rep] 2020 Nov 11; Vol. 10 (1), pp. 19562. Date of Electronic Publication: 2020 Nov 11.
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Typ publikacji:
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Journal Article; Research Support, Non-U.S. Gov't
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Język:
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English
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Imprint Name(s):
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Original Publication: London : Nature Publishing Group, copyright 2011-
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MeSH Terms:
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Green Fluorescent Proteins/*metabolism
Lactic Acid/*blood
Recombinant Proteins/*metabolism
Serine/*analysis
Animals ; Biosensing Techniques/methods ; Cells, Cultured ; Female ; Glycolysis ; Green Fluorescent Proteins/genetics ; HeLa Cells ; Humans ; Lactic Acid/metabolism ; Mice, Inbred ICR ; Molecular Imaging/methods ; Myocytes, Cardiac/drug effects ; Myocytes, Cardiac/metabolism ; Neurons/metabolism ; Oligomycins/pharmacology ; Pregnancy ; Pyruvic Acid/metabolism ; Recombinant Proteins/genetics
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References:
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Substance Nomenclature:
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0 (Oligomycins)
0 (Recombinant Proteins)
147336-22-9 (Green Fluorescent Proteins)
33X04XA5AT (Lactic Acid)
452VLY9402 (Serine)
8558G7RUTR (Pyruvic Acid)
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Entry Date(s):
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Date Created: 20201112 Date Completed: 20210422 Latest Revision: 20210422
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Update Code:
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20240105
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PubMed Central ID:
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PMC7659002
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DOI:
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10.1038/s41598-020-76440-4
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PMID:
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33177605
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Glycolysis is the metabolic pathway that converts glucose into pyruvate, whereas fermentation can then produce lactate from pyruvate. Here, we developed single fluorescent protein (FP)-based lactate and pyruvate indicators with low EC 50 for trace detection of metabolic molecules and live cell imaging and named them "Green Lindoblum" and "Green Pegassos," respectively. Green Lindoblum (EC 50 of 30 µM for lactate) and Green Pegassos (EC 50 of 70 µM for pyruvate) produced a 5.2- and 3.3-fold change in fluorescence intensity in response to lactate and pyruvate, respectively. Green Lindoblum measured lactate levels in mouse plasma, and Green Pegassos in combination with D-serine dehydratase successfully estimated D-serine levels released from mouse primary cultured neurons and astrocytes by measuring pyruvate level. Furthermore, live cell imaging analysis revealed their utility for dual-colour imaging, and the interplay between lactate, pyruvate, and Ca 2+ in human induced pluripotent stem cell-derived cardiomyocytes. Therefore, Green Lindoblum and Green Pegassos will be useful tools that detect specific molecules in clinical use and monitor the interplay of metabolites and other related molecules in diverse cell types.
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