Structural determinants of monohydroxylated bile acids to activate β1 subunit-containing BK channels

Anna Bukiya, Jacob McMilan, Abby L. Parrill, Alejandro Dopico

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Lithocholate (LC) (10-300 μM) in physiological solution is sensed by vascular myocyte large conductance, calcium- and voltage-gated potassium (BK) channel β1 accessory subunits, leading to channel activation and arterial dilation. However, the structural features in steroid and target that determine LC action are unknown. We tested LC and close analogs on BK channel (pore-forming cbv1+β1 subunits) activity using the product of the number of functional ion channels in the membrane patch (N) and the open channel probability (Po). LC (5β-cholanic acid-3α-ol), 5α-cholanic acid-3α-ol, and 5β-cholanic acid-3β-ol increased NPo (EC50 ∼45 μM). At maximal increase in NPo, LC increased NPo by 180%, whereas 5α-cholanic acid-3α-ol and 5β-cholanic acid-3β-ol raised NPo by 40%. Thus, the α-hydroxyl and the cis A-B ring junction are both required for robust channel potentiation. Lacking both features, 5α-cholanic acid-3β-ol and 5-cholenic acid-3β-ol were inactive. Three-dimensional structures show that only LC displays a bean shape with clear-cut convex and concave hemispheres; 5α-cholanic acid-3α-ol and 5β-cholanic acid-3β-ol partially matched LC shape, and 5α-cholanic acid-3β-ol and 5-cholenic acid-3β-ol did not. Increasing polarity in steroid rings (5β-cholanic acid-3α-sulfate) or reducing polarity in lateral chain (5β-cholanic acid 3α-ol methyl ester) rendered poorly active compounds, consistent with steroid insertion between β 1 and bilayer lipids, with the steroid-charged tail near the aqueous phase. Molecular dynamics identified two regions in β1 transmembrane domain 2 that meet unique requirements for bonding with the LC concave hemisphere, where the steroid functional groups are located.

Original languageEnglish (US)
Pages (from-to)2441-2451
Number of pages11
JournalJournal of lipid research
Volume49
Issue number11
DOIs
StatePublished - Dec 1 2008

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Large-Conductance Calcium-Activated Potassium Channels
Lithocholic Acid
Bile Acids and Salts
Steroids
cholanic acid
Voltage-Gated Potassium Channels
Lipid bilayers
Acids
Accessories
Lipid Bilayers
Molecular Dynamics Simulation
Ion Channels
Hydroxyl Radical
Muscle Cells
Functional groups
Sulfates
Blood Vessels
Molecular dynamics
Dilatation
Potassium

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Endocrinology
  • Cell Biology

Cite this

Structural determinants of monohydroxylated bile acids to activate β1 subunit-containing BK channels. / Bukiya, Anna; McMilan, Jacob; Parrill, Abby L.; Dopico, Alejandro.

In: Journal of lipid research, Vol. 49, No. 11, 01.12.2008, p. 2441-2451.

Research output: Contribution to journalArticle

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abstract = "Lithocholate (LC) (10-300 μM) in physiological solution is sensed by vascular myocyte large conductance, calcium- and voltage-gated potassium (BK) channel β1 accessory subunits, leading to channel activation and arterial dilation. However, the structural features in steroid and target that determine LC action are unknown. We tested LC and close analogs on BK channel (pore-forming cbv1+β1 subunits) activity using the product of the number of functional ion channels in the membrane patch (N) and the open channel probability (Po). LC (5β-cholanic acid-3α-ol), 5α-cholanic acid-3α-ol, and 5β-cholanic acid-3β-ol increased NPo (EC50 ∼45 μM). At maximal increase in NPo, LC increased NPo by 180{\%}, whereas 5α-cholanic acid-3α-ol and 5β-cholanic acid-3β-ol raised NPo by 40{\%}. Thus, the α-hydroxyl and the cis A-B ring junction are both required for robust channel potentiation. Lacking both features, 5α-cholanic acid-3β-ol and 5-cholenic acid-3β-ol were inactive. Three-dimensional structures show that only LC displays a bean shape with clear-cut convex and concave hemispheres; 5α-cholanic acid-3α-ol and 5β-cholanic acid-3β-ol partially matched LC shape, and 5α-cholanic acid-3β-ol and 5-cholenic acid-3β-ol did not. Increasing polarity in steroid rings (5β-cholanic acid-3α-sulfate) or reducing polarity in lateral chain (5β-cholanic acid 3α-ol methyl ester) rendered poorly active compounds, consistent with steroid insertion between β 1 and bilayer lipids, with the steroid-charged tail near the aqueous phase. Molecular dynamics identified two regions in β1 transmembrane domain 2 that meet unique requirements for bonding with the LC concave hemisphere, where the steroid functional groups are located.",
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N2 - Lithocholate (LC) (10-300 μM) in physiological solution is sensed by vascular myocyte large conductance, calcium- and voltage-gated potassium (BK) channel β1 accessory subunits, leading to channel activation and arterial dilation. However, the structural features in steroid and target that determine LC action are unknown. We tested LC and close analogs on BK channel (pore-forming cbv1+β1 subunits) activity using the product of the number of functional ion channels in the membrane patch (N) and the open channel probability (Po). LC (5β-cholanic acid-3α-ol), 5α-cholanic acid-3α-ol, and 5β-cholanic acid-3β-ol increased NPo (EC50 ∼45 μM). At maximal increase in NPo, LC increased NPo by 180%, whereas 5α-cholanic acid-3α-ol and 5β-cholanic acid-3β-ol raised NPo by 40%. Thus, the α-hydroxyl and the cis A-B ring junction are both required for robust channel potentiation. Lacking both features, 5α-cholanic acid-3β-ol and 5-cholenic acid-3β-ol were inactive. Three-dimensional structures show that only LC displays a bean shape with clear-cut convex and concave hemispheres; 5α-cholanic acid-3α-ol and 5β-cholanic acid-3β-ol partially matched LC shape, and 5α-cholanic acid-3β-ol and 5-cholenic acid-3β-ol did not. Increasing polarity in steroid rings (5β-cholanic acid-3α-sulfate) or reducing polarity in lateral chain (5β-cholanic acid 3α-ol methyl ester) rendered poorly active compounds, consistent with steroid insertion between β 1 and bilayer lipids, with the steroid-charged tail near the aqueous phase. Molecular dynamics identified two regions in β1 transmembrane domain 2 that meet unique requirements for bonding with the LC concave hemisphere, where the steroid functional groups are located.

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