Steroid ring hydroxylation patterns govern cooperativity in human bile acid binding protein.
Human ileal bile acid binding protein (I-BABP) is a member of the intracellular lipid binding protein family. This protein is thought to function in the transcellular transport and enterohepatic circulation of bile salts. Human I-BABP binds two molecules of glycocholate, the physiologically most abundant bile salt, with modest intrinsic affinity but a remarkably high degree of positive cooperativity. Here we report a calorimetric analysis for the binding of a broad panel of bile salts to human I-BABP. The interaction of I-BABP with nine physiologically relevant derivatives of cholic acid, chenodeoxycholic acid, and deoxycholic acid in their conjugated (glycine and taurine) and unconjugated forms was monitored by isothermal titration calorimetry. All bile salts bound to I-BABP with a 2:1 stoichiometry and similar overall affinity, but the derivatives of cholic acid displayed much higher Hill coefficients, a measure of macroscopic positive cooperativity. To test whether the cooperativity was dependent on individual structural features of the bile salt side chain, a series of side-chain-extended bile salts that lacked a hydrogen bond donor or acceptor at C-24 were chemically synthesized. These synthetic variants exhibited the same energetic and cooperativity profile as the naturally occurring bile salts. Our findings indicate that cooperativity in bile salt-I-BABP recognition is governed by the pattern of steroid B- and C-ring hydroxylation and not the presence or type of side-chain conjugation.
Tochtrop
Gregory P
GP
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, 660 South Euclid Avenue, Campus Box 8231, St. Louis, Missouri 63110, USA.
Bruns
Jamie L
JL
Tang
Changguo
C
Covey
Douglas F
DF
Cistola
David P
DP
D001665:Binding Sites; D002151:Calorimetry; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D002635:Chenodeoxycholic Acid / Q000737:chemistry / Q000378:metabolism; D019826:Cholic Acid / Q000737:chemistry* / Q000378:metabolism; D003840:Deoxycholic Acid / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D006900:Hydroxylation; D006913:Hydroxysteroid Dehydrogenases; D008024:Ligands; D008562:Membrane Glycoproteins; D011485:Protein Binding; D013696:Temperature; D013816:Thermodynamics; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)
D002352:Carrier Proteins; D002635:Chenodeoxycholic Acid; D019826:Cholic Acid; D003840:Deoxycholic Acid; D006913:Hydroxysteroid Dehydrogenases; D008024:Ligands; D008562:Membrane Glycoproteins; C037643:bile acid binding proteins; C435496:AKR1C2 protein, human; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)