Effect of the NBD-group position on interaction of fluorescently-labeled cholesterol analogues with human steroidogenic acute regulatory protein STARD1

Tugaeva, Kristina V.; Faletrov, Yaroslav V.; Allakhverdiev, Elvin S.; Shkumatov, V. M.; Maksimov, Eugene G.; Sluchanko, Nikolai N.

doi:10.1016/j.bbrc.2018.02.014

Cited by 12 publications

(8 citation statements)

References 47 publications

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“…Although the presence of the fast component indicates that the 22NC fluorophore can rotate with a rate similar for the free dye, we assigned this component to 22NC accommodated within the cholesterol‐binding cavity of STARD1, due to the following reasons. First, given the nanomolar ligand‐binding affinity [40] and the concentrations used (1 µ m ligand and 10 µ m pSTARD1), all 22NC molecules are expected to be bound to pSTARD1. Second, the solvatochromic property of 22NC would mean ~ 116 times lower fluorescence signal from the dissociated dye that could not be detected in the presence of the dominating protein‐bound form.…”

Section: Resultsmentioning

confidence: 99%

“…3). As we showed previously, in solution STARD1 66–285 predominantly exists in a monomeric conformation equivalent to the crystallographic monomer (PDB http://3P0L) [53] and is capable of binding steroids [26,40]. Likewise, we produced and characterized the START domain of the closest STARD1 homolog, STARD3, which, in the context of nonexpressed STARD1, is responsible for steroidogenesis in placenta [11,22].…”

Section: Resultsmentioning

confidence: 99%

“…In contrast to the phosphorylatable wild‐type transgene, STARD1 with the S195A amino acid substitution failed to rescue STARD1 −/− phenotype in knockout mice, characterized by intracellular accumulation of cholesterol droplets, low level of steroid hormones and neonatal lethality [9]. These effects of Ser195 phosphorylation lacked mechanistic explanation, as modification of Ser195 did not change STARD1 folding, cholesterol‐binding ability [28,40] or the mitochondrial import efficiency in vitro [32]. The functional role of STARD1 phosphorylation remains largely unclear but likely involves the regulation of STARD1 interaction with other proteins.…”

Section: Introductionmentioning

confidence: 99%

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Molecular basis for the recognition of steroidogenic acute regulatory protein by the 14‐3‐3 protein family

et al. 2020

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The human STARD1 protein supplies cholesterol to the mitochondria to initiate steroidogenesis. STARD1 functioning is regulated by binding to the 14‐3‐3 protein. We show that recognition by 14‐3‐3 is triggered by STARD1 phosphorylation at Ser57 and Ser195 by protein kinase A. Our observations are substantiated by biochemical evidence and crystal structures of 14‐3‐3 complexes with Ser57 and Ser195 phosphopeptides, revealing distinct roles for the two phosphosites in steroidogenesis regulation.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Molecular basis for the recognition of steroidogenic acute regulatory protein by the 14‐3‐3 protein family

et al. 2020

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“…To assess thermally-induced changes in FRP oligomeric state, we analyzed changes in their intrinsic Trp fluorescence (excitation at 297 nm; emission at 382 nm; slit width 5 nm, detector voltage 700 V) upon heating of 1 µM protein samples prepared in buffer F at a constant rate of 1 °C min −1 on a Cary Eclipse spectrofluorometer (Varian) equipped with a multicell holder and a Peltier temperature controller. The raw temperature dependencies, showing a single thermal transition, were transformed into dependences of completeness of transition on temperature 52 , 53 by linear approximation of the regions before and after the transition and representation of the data as percentage of the transition from the folded to the unfolded state. From these transformations, half-transition temperatures ( T 0.5 ) were directly determined.…”

Section: Methodsmentioning

confidence: 99%

OCP–FRP protein complex topologies suggest a mechanism for controlling high light tolerance in cyanobacteria

et al. 2018

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In cyanobacteria, high light photoactivates the orange carotenoid protein (OCP) that binds to antennae complexes, dissipating energy and preventing the destruction of the photosynthetic apparatus. At low light, OCP is efficiently deactivated by a poorly understood action of the dimeric fluorescence recovery protein (FRP). Here, we engineer FRP variants with defined oligomeric states and scrutinize their functional interaction with OCP. Complemented by disulfide trapping and chemical crosslinking, structural analysis in solution reveals the topology of metastable complexes of OCP and the FRP scaffold with different stoichiometries. Unable to tightly bind monomeric FRP, photoactivated OCP recruits dimeric FRP, which subsequently monomerizes giving 1:1 complexes. This could be facilitated by a transient OCP–2FRP–OCP complex formed via the two FRP head domains, significantly improving FRP efficiency at elevated OCP levels. By identifying key molecular interfaces, our findings may inspire the design of optically triggered systems transducing light signals into protein–protein interactions.

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“…Это позволяет рассматривать другие молекулы, содержащие такие фрагменты, в качестве перспективных кандидатов на роль соединений-зондов. В частности, такими соединениями являются производные стероидов, содержащие NBD-, или индольный фрагмент в боковой цепи [9].…”

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<i>In silico</i> analysis of interaction of compounds, containing photoactivatable groups,with human CYP7 enzymes

Диченко¹,

Хорецкий²,

Фалетров³

et al. 2020

Dokl. Akad. nauk

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In silico analysis of “protein-ligand” complexes of human CYP7 enzymes with modified borondipyrrome-tene (BODIPY) and steroids, containing photo-activated crosslinking groups, wasperformed in order to identify structural peculiarities of their interaction. It was found that BODIPY molecules and DHEA derivative with diazirine group are able to bind tightly with human steroid-hydroxylases. Binding affinity is comparable with corresponding values for essential ligands of the enzymes. Binding mode of the modified steroid corresponds to the binding mode of essential CYP7 ligands, so formation of hydroxylated products is possible. It was found that presence of both diazirine and NBD groups in a molecule significantly increases affinity of the compound in case of CYP7A1 and, especially, CYP7B1. Amino acid residues, located in a close proximity with photo-activated groups were detected, that can form covalent adducts with them. The obtained results can shed light on the mechanism of interaction of the compounds with recombinant human CYP7 enzymes in vitro. The results can also be used for the identification of modified amino acids of the proteins that are formed under photoactivation of the compounds in vitro.

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Effect of the NBD-group position on interaction of fluorescently-labeled cholesterol analogues with human steroidogenic acute regulatory protein STARD1

Cited by 12 publications

References 47 publications

Molecular basis for the recognition of steroidogenic acute regulatory protein by the 14‐3‐3 protein family

Molecular basis for the recognition of steroidogenic acute regulatory protein by the 14‐3‐3 protein family

OCP–FRP protein complex topologies suggest a mechanism for controlling high light tolerance in cyanobacteria

<i>In silico</i> analysis of interaction of compounds, containing photoactivatable groups,with human CYP7 enzymes

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