Litcius/Paper detail

GEM-IL: A highly responsive fluorescent lactate indicator

Ramsey Bekdash, Jose R. Quejada, Shunnosuke Ueno, Fuun Kawano, Kumi Morikawa, Alison D. Klein, Kenji Matsumoto, Tetz C. Lee, Koki Nakanishi, Amy Chalan, Teresa M. Lee, Rui Liu, Shunichi Homma, Chyuan‐Sheng Lin, Maria V. Yelshanskaya, Alexander I. Sobolevsky, Keisuke Goda, Masayuki Yazawa

2021Cell Reports Methods26 citationsDOIOpen Access PDF

Abstract

Lactate metabolism has been shown to have increasingly important implications in cellular functions as well as in the development and pathophysiology of disease. The various roles as a signaling molecule and metabolite have led to interest in establishing a new method to detect lactate changes in live cells. Here we report our development of a genetically encoded metabolic indicator specifically for probing lactate (GEM-IL) based on superfolder fluorescent proteins and mutagenesis. With improvements in its design, specificity, and sensitivity, GEM-IL allows new applications compared with the previous lactate indicators, Laconic and Green Lindoblum. We demonstrate the functionality of GEM-IL to detect differences in lactate changes in human oncogenic neural progenitor cells and mouse primary ventricular myocytes. The development and application of GEM-IL show promise for enhancing our understanding of lactate dynamics and roles.

Topics & Concepts

MetaboliteCell biologyFluorescenceBiologyComputational biologyMutagenesisFluorescent proteinGreen fluorescent proteinBiochemistryChemistryMutationGeneQuantum mechanicsPhysicsProtein Degradation and InhibitorsMitochondrial Function and PathologyATP Synthase and ATPases Research