A method was developed for measuring in vivo DNA synthesis after exposure to epidermal growth factor (EGF) or fibroblast growth factor (FGF) in a local area of mouse skin using ring-shaped forceps in combination with autoradiography. The technique should be useful for analysing the effects of growth factors on individual cells of the skin in vivo. EGF induced semisynchronized DNA synthesis in basal cells of the epidermis dose-dependently, but FGF did not. Time course study showed that EGF-induced DNA synthesis in basal cells increased with time for 24 h, and then decreased rapidly. EGF-induced DNA synthesis in basal cells was proportional to the time exposed to EGF (0-60 min). FGF and EGF both had little effect on dermal fibroblastic cells. The discrepancy between in vivo observations and those with cultured mammalian cells is discussed. 相似文献
Visible absorption and CD spectral and potentiometric studies on the His- and Tyr-containing ternary copper(II) complexes Cu(A)(L-B), where A refers to L-His, D-His, or L-Tyr and B to Lys, Tyr, Trp, Phe, Ala, Val, Arg, Glu, Asn, Gln, Ser, or Thr, were made to study ligand-ligand interactions in the complexes. While the CD spectral magnitudes in the d—d region are additive in the absence of side chain interactions and can be estimated from the magnitudes for the ternary systems involving DL-A or DL-B, deviation from the additivity was observed for Cu(L-His)(L-B) (B = LysH, Tyr, Trp, or Phe) and Cu(L-Tyr)(L-Trp). From the stability constants determined at 25 °C and I = 0.1 M (KNO3), the equilibrium constants, K, for the following hypothetical equilibria were calculated to be large (0.14–0.60) for formation of Cu(L-/D-His)(L-B)(B = Tyr or Trp) and Cu(D-His)(L-Phe) with Cu(en)(L-Ala) as standard: The positive values indicate the stabilization due to the stacking between the imidazole ring of His and the aromatic side chain of L-B. Solvent dependence of the CD spectra for Cu(L-His)(L-LysH) and Cu(L-His) L-Trp) further supported the existence of the intramolecular electrostatic and hydrophobic interactions. 相似文献
The axial ligations of nitrogenous bases to the five-coordinate chloro-meso-tetraphenylporphyrinatochromium(III) [Cr(III)(TPP)(Cl)] were studied in a non-coordinating solvent, dichloromethane (CH2Cl2), by spectrophotometric methods. A correlation exists between log K for the axial ligation: and pKa for the N-donor ligand. This correlation suggests that ligand to metal σ bonding contributes to the complex formation, rather than does metal to ligand π back-donation. 相似文献
Pyroglutamyl-lysyl-leucyl-argininal (Pyr-Lys-Leu-Argal) immobilized on gel matrix through the epsilon-amino group of its lysine residue was shown to be an efficient biospecific affinity adsorbent for purification of urokinase. Pyr-Lys-Leu-Argal dibutylacetal, a precursor of this immobilized ligand, was synthesized by a fragment condensation procedure, in which one of the thermolysin-digestion products of leupeptin dibutylacetal, H-Leu-Argal dibutylacetal, was used as a key intermediate. The precursor was coupled to CH-Sepharose 4B with the aid of a water-soluble carbodiimide, and its acetal protecting group was then removed by mild acid treatment to free the essential aldehyde function. The Sepharose derivative thus prepared was shown to adsorb urokinase selectively and effectively from a crude human urine preparation at neutral pH and to release the bound enzyme under mild acidic conditions. The present technique afforded a highly purified urokinase preparation abundant in the high-molecular form with 90% recovery. The complex formed between urokinase and the immobilized ligand was found to have a dissociation constant of about 2 X 10(-4)M. 相似文献
The alkali light chain of rabbit skeletal muscle myosin, A1, was cyanylated with 2-nitro-5-thiocyanobenzoic acid, and the peptide bond at Cys 177 was subsequently cleaved in the presence of 0.05 M CaCl2. Two peptide fragments, from the N-terminal to the residue 176 (CF1) and from the residue 177 to the C-terminal (CF2), were obtained. The CD spectrum and the difference UV absorption spectrum induced by CaCl2 suggested that CF1 largely retained the higher order structure of A1. The CF1 fragment, however, could neither incorporate subfragment-1 (S-1) by an exchange reaction, nor bind with the renatured 20K fragment of S-1 heavy chain. On the other hand, the C-terminal fragment of 14 residues, CF2, could bind with the 20K fragment of S-1 heavy chain. These results indicate that the binding site of the alkali light chain for the heavy chain of myosin is located within the C-terminal 14 residues. 相似文献
The prolactin receptor localized in rabbit mammary gland membranes has been identified by affinity labelling using covalent cross-linking agents such as a unique protein chain of approximately 32,000 daltons. After partial purification (5,000-fold) of these receptors from mammary gland homogenate, polyclonal antibodies, which specifically and completely inhibit prolactin binding in all organs and in all species studied, were raised. These antibodies possessed prolactin-like biological activity (casein synthesis) on rabbit mammary gland explants. Monoclonal antibodies specifically directed against the binding domain of the receptor were also obtained. These antibodies were more species-specific than the polyclonal antibodies. The most potent (M110) possessed higher affinity than prolactin for the receptor and could be a very effective tool to elucidate the structure of the receptor and its immunological detection. 相似文献
Dictyosiphon foeniculaceus from Sweden and Newfoundland was studied in laboratory culture. Zoids from unilocular sporangia developed into dioecious microscopic filamentous gametophytes which produced uniseriate plurilocular gametangia in low temperatures (0 to 8 °C). Zygotes and unfused isogametes gave rise to filamentous protonemata on which parenchymatous macroscopic sporophytes were formed. Isolates from Sweden and Newfoundland were interfertile. Although formed in culture, genetically unisexual sporophytes were not detected in nature. Female gametes ofD. foeniculaceus produced a sexual pheromone. It was identified as finavarrene, which is also known as the sperm attractant inAscophyllum nodosum. 相似文献
The binding of catechol estrogens (2-hydroxyestrone, 4-hydroxyestrone, 2-hydroxyestradiol, and 4-hydroxyestradiol) to estrogen receptors in 7,12-dimethylbenz(a)anthracene (DMBA)-induced rat mammary tumor cytosols was investigated. Cytosol estrogen receptors exhibited high affinities (Ka = 1.12-1.88 X 10(8) M-1) for all catechol estrogens as well as estradiol. The receptor level of catechol estrogens (46.1-97.5 fmol/mg protein) was 1.6-3.0 times higher than that of estradiol; especially the binding of 4-hydroxyestrone to estrogen receptors was the highest of all catechol estrogens and estradiol. In judging the receptor level of more than 20 fmol/mg protein to be positive, the binding of catechol estrogens to estrogen receptors was approximately correlated with that of estradiol. The positive receptor level of catechol estrogens was found in a half of tumor cytosols which showed the negative receptor level of estradiol. These results suggested that characteristic estrogen receptors indicating high affinities for catechol estrogens might be present in rat mammary tumor cytosols. 相似文献
Summary An investigation of the removal and recovery of urnnium from aqueous systems using microbial biomass has been described previously (Nakajima et al. 1982). To establish which microorganisms accumulate the most uranium, we extended our investigation of uranium uptake to 83 species of microorganisms, 32 bacteria, 15 yeasts, 16 fungi and 20 actinomycetes. Of these 83 species of microorganisms tested, extremely high uranium-absorbing ability was found in Pseudomonas stutzeri, Neurospora sitophila, Streptomyces albus and Streptomyces viridochromogenes.The selective accumulation of heavy metal ions by various microorganisms has also been examined. Uranyl, mercury and lead ions were readily accumulated by almost all the species of microorganisms tested. Actinomycetes and fungi differ from many bacteria and most yeasts in their selective accumulation of uranium and mercury.In addition to this fundamental research, uranium recovery was investigated in immobilized Streptomyces albus, a microorganism with high uranium-uptake ability. These immobilized cells adsorbed uranium readily and selectively. The immobilized cells recovered uranium almost quantitatively and almost all uranium absorbed was desorbed with 0.1 M Na2CO3. The dry weight of the free cells decreased by 50% during 5 adsorption-desorption cycles. However, the dry weight of the immobilized cells decreased by only 2% during 5 cycles. These results showed that microbial cells are more stable after immobilization and can be used repeatedly for the process of uranium adsorption-desorption. 相似文献