Fumito Tani

Opioid antagonist peptides derived from κ-casein

Opioid antagonists were sought in the fragments of kappa-casein which were obtained by chemical synthesis and enzymic digestion. A synthetic bovine kappa-casein peptide (35-41), Tyr-Pro-Ser-Tyr-Gly-Leu-Asn (casoxin A) showed opioid antagonist activity at 200 microM in the guinea pig ileum assay. A synthetic peptide Tyr-Pro-Tyr-Tyr (casoxin B) which is found in bovine and human kappa-casein, also showed opioid antagonist activity at 100 microM. Another opioid antagonist peptide (casoxin C) was isolated from tryptic digests of bovine kappa-casein by reverse-phase HPLC. The structure of the peptide was Tyr-Ile-Pro-Ile-Gln-Tyr-Val-Leu-Ser-Arg, which corresponded to kappa-casein (25-34). Casoxin C was active at 5 microM in the guinea pig ileum assay. Thus, bovine kappa-casein contains three potential opioid antagonist sequences.

Supramolecular Structures and Photoelectronic Properties of the Inclusion Complex of a Cyclic Free‐Base Porphyrin Dimer and C60

A cyclic free-base porphyrin dimer H4-CPD(Py) (CPD = cyclic porphyrin dimer) linked by butadiyne moieties bearing 4-pyridyl groups self-assembles to form a novel porphyrin nanotube in the crystalline state. The cyclic molecules link together through nonclassical C-H⋅⋅⋅N hydrogen bonds and π–π interactions of the pyridyl groups along the crystallographic a axis. H4-CPD(Py) includes a C60 molecule in its cavity in solution. In the crystal structure of the inclusion complex (C60⊂H4-CPD(Py)), the dimer “bites” a C60 molecule by tilting the porphyrin rings with respect to each other, and there are strong π–π interactions between the porphyrin rings and C60. The included C60 molecules form a zigzag chain along the crystallographic b axis through van der Waals contacts with each other. Femtosecond laser flash photolysis of C60⊂H4-CPD(Py) in the solid state with photoexcitation at 420 nm shows the formation of a completely charge-separated state {H4-CPD(Py)·+ + C60·−}, which decays with a lifetime of 470 ps to the ground state. The charge-carrier mobility of the single crystal of C60⊂H4-CPD(Py) was determined by flash photolysis time-resolved microwave conductivity (FP-TRMC) measurements. C60⊂H4-CPD(Py) has an anisotropic charge mobility (Σμ = 0.16 and 0.13 cm2 V(−1)  s(−1)) along the zigzag chain of C60 (which runs at 45° and parallel to the crystallographic b axis). To construct a photoelectrochemical cell, C60⊂H4-CPD(Py) was deposited onto nanostructured SnO2 films on a transparent electrode. The solar cell exhibited photovoltaic activity with an incident photon to current conversion efficiency of 17%.

Allosteric binding of an Ag+ ion to cerium(IV) bis-porphyrinates enhances the rotational activity of porphyrin ligands

A series of cerium(IV) bisporphyrinate double-deckers [Ce(bbpp)2] (BBPP = 5,15-bis(4-butoxyphenyl) porphyrin dianion), [Ce(tmpp)2] (TMPP = 5,10,15,20-tetrakis(4-methoxyphenyl)-porphyrin dianion), [Ce(tfpp)2] (TFPP = 5,10,15,20-tetrakis(4-fluorophenyl)porphyrin dianion), [Ce(tmcpp)2] (TMCPP = 5,10,15,20-tetrakis(4-methoxycarbonylphenyl)porphyrin dianion), and [Ce(tmpp)(tmcpp)] was prepared. They bind three Ag+ ions to their concave porphyrin pi subunits (pi-clefts) according to a positive homotropic allosteric mechanism with Hill coefficients (nH) of 1.7-2.7. The rotation rates of the porphyrin ligands in [Ce(bbpp)2] were evaluated to be 200 s-1 at 20 degrees C (delta G++293 = 14.1 kcal mol-1) and 220 s-1 at -40 degrees C (delta G++233 = 11.0 kcal mol-1) without and with Ag+ ions, respectively. These results consistently support our unexpected finding that Ag+ binding can accelerate rotation of the porphyrin ligand. On the basis of UV-visible, 1H NMR, and resonance Raman spectral measurements, the rate enhancement of the rotational speed of the porphyrin ligands is attributed to conformational changes of the porphyrin in cerium(IV) bis-porphyrinate induced by binding of Ag+ guest ions in the clefts. This novel concept of positive homotropic allosterism is applicable to the molecular design of various supramolecular and switch-functionalized systems.

Opioid Peptides from Milk Proteins

Fragment peptides of milk proteins containing Tyr-X-Phe or Tyr-X1-X2-Phe were synthesized and their opioid activities were evaluated by the guinea pig ileum and radioreceptor assays. Among the peptides tested, the human β-casein(51 - 54) amide, Tyr-Pro-Phe-Val-NH2 (valmuceptin) was the most active, more than the bovine counterpart, morphiceptin. The human β-casein(41 - 44) amide, Tyr-Pro-Ser-Phe-NH2, and the α-lactalbumin (50-53) amide, Tyr-Gly-Leu-Phe-NH2, were also active in the guinea pig ileum assay. These were named β-casorphin and α-lactorphin, respectively. All of these peptides were not as active in the mouse and rabbit vas deferens assays.

Synthetic models for the active site of cytochrome P450

Abstract As cytochrome P450 models, two kinds of iron porphyrins were synthesized, each of which has an alkanethiolate axial ligand and hydroxyl groups inside the molecular cavities. The stable dioxygen adducts were obtained by the reaction of the ferric complexes with KO 2 under an oxygen atmosphere. This study first demonstrated a hydrogen bond to bound dioxygen in an oxy form of a thiolate-coordinated heme model system. These results are discussed in relation to the process of dioxygen binding and activation in cytochrome P450.

Characterization of β-Lactotensin, a Bioactive Peptide Derived from Bovine β-Lactoglobulin, as a Neurotensin Agonist

β-Lactotensin (β-LT: His-Ile-Arg-Leu) is an ileum-contracting peptide derived from residues No. 146-149 of bovine β-lactoglobulin. The ileum-contracting activity of β-LT was blocked by the NT1 antagonist SR48692. β-LT was selective for the neurotensin NT2 receptor while neurotensin was selective for the NT1 receptor. β-LT is the first natural ligand showing selectivity for the NT2 receptor. β-LT showed hypertensive activity after intravenous administration at a dose of 30 mg/kg in conscious rats, while neurotensin showed hypotensive activity. The hypertensive activity of β-LT was blocked by levocabastine (1 mg/kg, i.v.), an NT2 antagonist. SR48692, which blocked the hypotensive activity of neurotensin, had no effect on the hypertensive activity of β-LT. These results suggest that the hypertensive activity of β-LT is mediated by the NT2 receptor. It was concluded that the NT1 and NT2 receptors mediate the opposite effect on blood pressure.

Enantioselective oxidation of sulfides catalyzed by iron complexes of chiral “twin coronet” porphyrins

Abstract Ferric porphyrins bearing chiral binaphthalene moieties on their both faces catalyze asymmetric oxidation of sulfides with iodosobenzene as an oxidant in moderate to good optical and chemical yields. Enantiomeric excesses (e.e.) were markedly improved in 2–3 times by the addition of 1-methylimidazole. The mechanism of oxidation and chiral induction are discussed.

Heat-induced Transparent Gel Formation of Bovine Serum Albumin

The formation of transparent gels by 6% bovine serum albumin (BSA), pH 7.5, was examined by one- and two-step heating methods. Heating of the BSA solutions at various NaCl concentrations produced transparent gels at 25-50mM NaCl and transparent sols at 0-20 mM NaCl (one-step heating method). The transparent sol obtained by heating without NaCl was reheated after mixing with various amounts of NaCl (two-step heating method I). The result was almost identical to that obtained by the one-step heating method. However, when the first heating was done with 10 mM NaCl, transparent gels were obtained over a wide range of NaCl concentrations with a second heating (two-step heating method II). Analyses of sols obtained at various NaCl concentrations by gel permeation chromatography and transmission electron microscopy showed the presence of linear polymers in the heated BSA sol (10 mM NaCl) and gel networks formed by the linear polymers (20 mM NaCl). The mechanism of transparent gel formation in BSA may be similar to that in ovalbumin.

Identification of casoxin C, an ileum-contracting peptide derived from bovine κ-casein, as an agonist for C3a receptors

Casoxin C (Tyr-Ile-Pro-Ile-Gln-Tyr-Val-Leu-Ser-Arg) is a bioactive peptide that was isolated from a tryptic digest of bovine kappa-casein as an anti-opioid peptide in longitudinal strips of guinea pig ileum. Casoxin C also evokes contraction of the ileal strips, and we found that this process was biphasic with rapid and slow components. The contractile profile was very similar to that of human complement C3a(70-77), which is the COOH-terminal octapeptide of C3a and has, although less potent, qualitatively the same biological activities as C3a. Casoxin C also has homology with C3a(70-77). The rapid contraction was mediated by histamine release and the slow contraction was mediated by a prostaglandin E2-like substance, judging from the effects of various pharmacological inhibitors and antagonists on the ileal contraction. Casoxin C had affinity for C3a receptors (IC50 = 40 microM) in the radioreceptor assay. In addition, casoxin C showed phagocyte-stimulating activities. Casoxin C is therefore the first milk-derived peptide identified, that acts through complement C3a receptors.

Submillisecond-lived photoinduced charge separation in inclusion complexes composed of Li + @C60 and cyclic porphyrin dimers†

Lithium ion encapsulated [60]fullerene (Li+@C60) is included within a free base and nickel complex of a cyclic porphyrin dimer (M-CPDPy, M = H4 and Ni2) to afford supramolecules (Li+@C60⊂M-CPDPy) in a polar solvent (benzonitrile) with the association constants of 2.6 × 105 M−1 and 3.5 × 105 M−1, respectively. From the electrochemical analysis, the energies of the charge-separated (CS) states are estimated to be 1.07 eV for Li+@C60⊂H4-CPDPy and 1.20 eV for Li+@C60⊂Ni2-CPDPy. Both values are lower than the triplet excited energies of the fullerene and porphyrin. Upon the photoexcitation at the Q-band of the porphyrin chromophore of Li+@C60⊂H4-CPDPy, electron transfer from the triplet excited state of the free base porphyrin to Li+@C60 occurs to produce the CS state. Li+@C60⊂Ni2-CPDPy also undergoes photoinduced electron transfer to produce the CS state. The lifetimes of the resulting CS states are 0.50 ms for Li+@C60⊂H4-CPDPy and 0.67 ms for Li+@C60⊂Ni2-CPDPy. These remarkably long CS lifetimes are the best values ever reported for non-covalent porphyrin-fullerene supramolecules in solution and are attributable to the lower CS energies than the triplet energy of each chromophore.