GTP synthases. Proton pumping and phosphorylation in ligand-receptor-G alpha-protein complexes

• Published in: Receptors & channels Vol. 4; no. 2; p. 111
• Main Authors: Nederkoorn, P H, Timmerman, H, Donné-Op Den Kelder, G M, Timms, D, Wilkinson, A J, Kelly, D R, Broadley, K J, Davies, R H
• Format: Journal Article
• Language: English
• Published: England 1996
• Subjects: Amino Acid Sequence; GTP-Binding Proteins - chemistry; GTP-Binding Proteins - metabolism; Guanosine Triphosphate - metabolism; Ligases - metabolism; Molecular Sequence Data; Phosphorylation; Protein Conformation; Proton Pumps; Protons; Receptors, Adrenergic, beta-1 - metabolism
• ISSN: 1060-6823

A structural model for a ligand-receptor-Gs alpha-protein complex to function as a GTP synthase is presented. The mechanism which is dependent on the movement and rotation of the G alpha-protein alpha 2-helix is seen to involve the delivery of, at least, one proton to the phosphorylation site in the rotation of this helix. The cycle is driven by a ligand-mediated proton pump through the alpha-helices of the receptor, attachment of the conserved Tyr-Arg-Tyr receptor proton shuttle being made to an aspartate group on the Gs alpha-protein terminal sidechain, which is itself linked to the Asn-Gln interaction known to control movement and rotation of the alpha 2-helix between .GDP and .GTP structures. The energetics of proton transfer through the shuttle mechanism and delivery of a proton to the aspartate group are shown to be sufficient to rupture this controlling interaction and its associated backbone bond. The complex leads to full spatial and energetic definition of the receptor proton shuttle mechanism, while there is a striking association of further Tyrosine and Arginine residues in the vicinity of the Gs alpha-protein Asn-Gln interaction. Calculations at the HF 6-31G** level confirm that a critical balance between ion pair and neutral forms of Tyr-Arg interactions under multiply hydrogen bonded conditions in a hydrophobic environment controls proton transfer and recovery mechanisms. The intrinsic preference of the neutral Tyr-Arg form over the ion-pair is 14.0 kcal/mol. Activation of the Tyrosine oxygen atom in the neutral form by single-NH or -OH groups reduces this difference by some 6.4-8.6 kcal/mol but the dominance of the neutral form is maintained. The expected slight overestimates are consistent with the maximum activation enthalpy of 11.0-12.0 kcal/ mol required to initiate proton transfer through the shuttle. The extended form of the shuttle with the Arginine acting competitively between the two Tyrosine residues allows interpretation of observed enthalpic differences in ligand binding with and without the presence of GTP. The uniqueness of Gs proteins among the G alpha-proteins is seen as their inability to transfer a proton directly through the alpha 2-helix switch Asn-Gln residues. A possible proton pathway to the mid-point of the Gs alpha-protein alpha 2 helix is outlined.