TY - JOUR
T1 - Cardiolipin in the spotlight
T2 - Quantitative analysis and fluorescence-based competitive binding assay
AU - Dimitrijevs, Pavels
AU - Arsenyan, Pavel
N1 - Funding Information:
This work was supported by Latvian Institute of Organic Synthesis internal grant ( IG-2020-01 for Pavels Dimitrijevs) and SAM project Nr. 1.1.1.1/19/A/016. Cardiac, kidney and brain tissue samples of healthy C57Bl/6 J mice and cardiac tissue from healthy Sprague Dawley rats were kindly provided by LIOS Laboratory of Pharmaceutical Pharmacology. Authors would like to sincerely thank Dr. S. Belyakov for X-ray analysis, K. Leduskrasts for photo-physical experiments and I. Domracheva for culturing the cells.
Publisher Copyright:
© 2021 Elsevier B.V.
PY - 2021/11/1
Y1 - 2021/11/1
N2 - Cardiolipin (CL) is a key phospholipid responsible for mitochondrial function and cristae integrity. The CL level is associated with various diseases characterized by mitochondrial dysfunction, including ischemic heart diseases and cancer. CL is an attractive target for mitochondria-specific drugs, but unnecessary interaction with CL might lead to detrimental side effects such as heart failure and kidney dysfunction. Thus, a simple and robust method for CL quantification and a reliable assay for the determination of drug affinity for CL are desired. We report a new fluorescent CL-specific probe with impressive photophysical properties that allows CL quantification in mitochondrial fractions isolated from cell and tissue homogenates and enables estimates of drug affinity for CL in the first fluorescence-based competitive binding assay. It was found that CL concentration is elevated in mitochondrial fractions isolated from cancer cells and cells with high proliferation rate (up to 108.5 ± 16.0 nmol/mg prot in mouse colon carcinoma cells, CT-26). CL concentration in mitochondria from brain tissue (66.11 ± 5.78 nmol/mg prot) is circa twice higher than in heart and kidney mitochondria (37.49 ± 8.69 and 33.95 ± 5.32 nmol/mg prot, respectively.) Generally, positively charged substances bind with CL, but their affinity is highly variable with EC50 values ranging from sub-micromolar to millimolar concentration.
AB - Cardiolipin (CL) is a key phospholipid responsible for mitochondrial function and cristae integrity. The CL level is associated with various diseases characterized by mitochondrial dysfunction, including ischemic heart diseases and cancer. CL is an attractive target for mitochondria-specific drugs, but unnecessary interaction with CL might lead to detrimental side effects such as heart failure and kidney dysfunction. Thus, a simple and robust method for CL quantification and a reliable assay for the determination of drug affinity for CL are desired. We report a new fluorescent CL-specific probe with impressive photophysical properties that allows CL quantification in mitochondrial fractions isolated from cell and tissue homogenates and enables estimates of drug affinity for CL in the first fluorescence-based competitive binding assay. It was found that CL concentration is elevated in mitochondrial fractions isolated from cancer cells and cells with high proliferation rate (up to 108.5 ± 16.0 nmol/mg prot in mouse colon carcinoma cells, CT-26). CL concentration in mitochondria from brain tissue (66.11 ± 5.78 nmol/mg prot) is circa twice higher than in heart and kidney mitochondria (37.49 ± 8.69 and 33.95 ± 5.32 nmol/mg prot, respectively.) Generally, positively charged substances bind with CL, but their affinity is highly variable with EC50 values ranging from sub-micromolar to millimolar concentration.
KW - Cardiolipin
KW - Competitive binding
KW - Fluorescent probe
KW - Quantitative analysis
UR - http://www.scopus.com/inward/record.url?scp=85111897765&partnerID=8YFLogxK
U2 - 10.1016/j.snb.2021.130537
DO - 10.1016/j.snb.2021.130537
M3 - Article
AN - SCOPUS:85111897765
SN - 0925-4005
VL - 346
JO - Sensors and Actuators B: Chemical
JF - Sensors and Actuators B: Chemical
M1 - 130537
ER -
