Journal
ENDOCRINOLOGY
Volume 142, Issue 1, Pages 502-505Publisher
ENDOCRINE SOC
DOI: 10.1210/en.142.1.502
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Funding
- NICHD NIH HHS [R01 HD23236, T32 HD07031] Funding Source: Medline
- EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH &HUMAN DEVELOPMENT [T32HD007031, R01HD023236] Funding Source: NIH RePORTER
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Steroidogenic acute regulatory protein (StAR) and peripheral-type benzodiazepine receptor (PBR) have both been implicated in the transport of cholesterol across mitochondrial membranes in steroidogenic cells. Therefore, we hypothesized that StAR and PER were associated in this process. To test this hypothesis, we measured fluorescence energy transfer (FET) between these proteins by fusing enhanced green fluorescent protein (GFP, donor fluorophore) and yellow fluorescent protein (YFP, acceptor fluorophore) to the C-taminus of ovine StAR (37GFP) and ovine PER (PBRYFP), respectively. These intrinsically fluorescent proteins were stably transfected into Cos-7 cells and determined to be biologically active. For FET to occur the appropriate fluorescent molecules need to be < 100 from each other. We observed 22.0 +/- 0.9 % energy transfer efficiency for 37GFP and PBRYFP, a 4.9 fold increase above non-specific energy transfer between free GFP and PBRYFP (p < .0001). Thus, it appears that StAR and PER are closely associated in mitochondrial membranes and that these molecules may interact in the transportation of cholesterol.
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