Conformations and conformational interconversions of diastereomeric cyclic tetraprolines.

Cyclic tetrapeptides exclusively composed of L- and D-Pro have been studied by theoretical means (conformational searches and molecular mechanics calculations using the CHARMM program) supported by 1H-NMR spectroscopy, X-ray analysis and chiroptical measurements. We explored the entire conformational space of the diastereomers cyclo(LLLL-Pro4) (I), cyclo(LDLD-Pro4) (II) and cyclo(LLDD-Pro4) (III) including the low-energy conformations and the related interconversion paths. The conformational interconversions were found to be restricted to cis/trans isomerisations of the amide bonds. Owing to the polycyclic nature of cyclo(Pro4) most of the cis/trans transitions are hindered by energy barriers higher than 30 kcal/mol (up to 150-200 kcal/mol). A few transitions are characterized by computed energy barriers comparable to those found in linear -Xxx-Pro- sequences (approximately 18 kcal/mol), and are therefore experimentally significant. Experimental evidence has been obtained in the case of cyclo(LDLD-Pro4), where two enantiomers are interconverted by a series of 4 cis/trans isomerisations ctct-->cttt-->tttt-->tctt-->tctc. The Erying activation parameters of this reaction were determined in H2O and in DMF by chiroptical measurements (delta H not equal to = 44 and 28 kcal/mol; delta S not equal to = 59 and 22 cal K-1 mol-1, respectively), and correlated with the calculated barriers. In I and III comparable series of four cis/trans isomerisations relate two main conformations with the peptide bond sequences ctct and tctc. In compound I pseudorotational images are interconverted via ctct-->ccct-->cctt-->cctc-->tctc.(ABSTRACT TRUNCATED AT 250 WORDS)

Suppression of human chorionic gonadotropin in the human placenta at term by human thyroid-stimulating hormone in vitro.

Human chorionic gonadotropin (hCG) exerts a clinically apparent negative feedback on the secretion of human thyroid-stimulating hormone (hTSH) in pregnancy, and the two have cross-reactivity for the TSH receptor in membrane preparations of the thyroid. We examined whether hTSH, in turn, has an influence on the secretion and synthesis of hCG in short-term cultures of human placenta at term. A dose- and time-dependent decrease in the extracellular hCG concentration caused by hTSH was demonstrated. To examine whether hTSH inhibits de novo synthesis of hCG or decreases hCG depletion, we determined the amount of hCG secreted and the size of the intracellular pool by using an enzyme immunoassay. By incorporating a radiolabeled amino acid in the hCG molecule, we measured the amount of hCG synthesized de novo. We concluded that hTSH acts by decreasing the rate of de novo synthesis of placental hCG.

Conformation and formation tendency of the cyclotetrapeptide cyclo (D-Pro-D-Pro-L-Pro-L-Pro): experimental results and molecular modeling studies.

The title compound represents the smallest member of cyclic proline peptides corresponding to the general formula c(DDLL-Pro4)n with a strictly D,D,L,L double-alternating sequence of the chiral amino acid residues. The cyclopeptides with n greater than or equal to 2 could be synthesized from both DDLL-Pro4 (1) and DLLD-Pro4 (2). The cyclic monomer (n = 1) resulted only from 2, whereas not even a trace could be found by cyclization of 1. The peptide exists in a strongly strained Ci symmetrical conformation (x-ray analysis) with alternating cis and trans peptide bonds (ctct form I). The cis peptide bonds deviate from planarity (omega = 22 degrees); two of the pyrrolidine rings show a "South" conformation (phi = -94 degrees), whereas the other residues exhibit C alpha-endo puckering (phi = -124 degrees). Two of the psi angles surprisingly occur at +41 degrees (anti-cis'), the others are located in the trans' region. A quantitative ring opening occurs with trifluoroacetic acid at room temperature. In solution the existence of an isomeric ctcc sequence (form Ia) is indicated. Dreiding model studies also suggested a favorable conformation with a tctc sequence (form II). Consequently, we performed molecular mechanics calculations, based on the CHARMM force field and semiempirical quantum mechanical AM1 calculations (MOPAC program). Pronounced differences in the backbone parameters were found using these two methods. However, the theoretical studies evidenced the experimentally obtained differences in the cyclization tendencies of the linear precursors.