Differential effects of zinc on functionally distinct human growth hormone mutations.
Human growth hormone (hGH) binds and activates lactogenic receptors by a sequential receptor dimerization mechanism. The affinity for the first lactogenic receptor is increased due to one zinc molecule linking hGH residues H18 and E174, located in helices 1 and 4, respectively, with two adjacent residues in the lactogenic receptor (D187 and H188). Two functionally unique groups of mutant hGHs have been identified. Addition of 25 microM zinc to lactogenic bioassays differentially affects mutant activities based on which group they belong to. One mutation (G120R) is located within the binding surface of hGH that interacts with the second lactogenic receptor. In the presence of endogenous zinc, G120R reduces the maximal activity of hGH without altering either the agonist or antagonist phases of the bell-shaped dose-response curve. Addition of zinc to this assay further reduces the activity of this protein. In contrast, mutations within a hydrophobic motif in hGH that functionally couples the two lactogenic receptor binding sites decrease the sensitivity (right-shift) of the agonist phase of the dose-response curve without similarly affecting the antagonist phase. The addition of zinc to these lactogenic assays increases the sensitivity (left-shifts) of the dose-response curves, largely negating the effect of these mutations. The effects of zinc differentiate between mutations within these two distinct functional motifs by limiting the pool of potential conformations that are available for binding within either of the receptor binding sites of this ligand.
Duda
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KM
Ohio State Biochemistry Program, The Ohio State University, 1925 Coffey Road, Columbus, OH 43210, USA.
Brooks
C L
CL
D019943:Amino Acid Substitution; D001681:Biological Assay; D004305:Dose-Response Relationship, Drug; D019382:Human Growth Hormone / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D009154:Mutation; D015032:Zinc / Q000378:metabolism*
D019382:Human Growth Hormone; D015032:Zinc