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1.
Development of effective resuscitation agents for blood-loss replacement in trauma or surgery is extremely important despite substantial improvements in screening methods of blood from human donors. This paper reports the design and synthesis of peptides that mimic the natural environment of the heme group in myoglobin (Mb) and in the - and -subunits of human adult hemoglobin (Hb). The designs were based on the fact that the heme group in the aforementioned proteins is sandwiched between helices E and F. Fifteen test peptides and six control peptides were synthesized, and their ability to form stable complexes with heme was investigated. It was found that none of the control peptides or proteins was able to bind heme. However, each of the peptides that were designed to mimic the E--F helices, and even shorter designs, which removed from this region residues that do not contribute to contacts with the heme group, were each able to bind one mole of heme per mole of peptide forming peptide–heme complexes that were stable to manipulation and behaved as single molecular species. Oxygen binding measurements on the reduced peptide–heme complexes showed that these compounds bind oxygen and give visible spectra that were typical of oxygenated heme-proteins. In oxygen binding measurements done under different partial pressures of oxygen, the heme–peptide complexes gave hyperbolic oxygen-saturation curves, but showed slight differences in their P50 values. The P50 values ranged from 3.8 mmHg for the heme–peptide B7 complex to 13.7 mmHg for the heme–peptide D13 complex (under the same conditions, P50 values for Hb and Mb were 34.0 and 5.5 mmHg, respectively). It is concluded that peptide constructs designed to mimic the heme-binding regions of Mb or the Hb subunits were able to form coordinate 1:1 complexes with heme, and these complexes bind oxygen in a manner expected for single subunit heme proteins. 相似文献
2.
Quantitative measurement of cell-bubble attachment in bubble columns using foam flotation techniques
A simple and convenient system for quantitatively measuring the number of adsorbed animal cells per unit of bubble surface area (, unit: cells/cm2) was developed. The system was successfully applied to recombinant Chinese hamster ovary (r-CHO) suspension cultures to investigate the dynamic cell-bubble attachment in a bubble column. In serum-free medium, values increased with bubble rising height (H) and cell concentration (C) and then became constant (about 1750 cells/cm2) when H and C were sufficiently high. In medium containing protective additives, the trends of values with H were similar to that in serum-free medium. Compared with serum-free medium, polyvinyl-pyrrolidone (PVP) increased the values to 1941 cell/cm2 whereas other tested additives decreased the values of in some different degree. 相似文献
3.
Induction of petite mutation and inhibition of synthesis of respiratory enzymes in various yeasts 总被引:10,自引:0,他引:10
C. J. E. A. Bulder 《Antonie van Leeuwenhoek》1964,30(1):1-9
A systematic investigation covering a wide diversity of yeast species was made on the appearance of respiratory deficient (petite) mutants after treatment with acriflavine. Petite mutants were obtained from certain species only, but in these species all strains were found to have in common the property of giving rise to petite mutants; such species were designated as petite positive. Species failing to give rise to petite mutants were accordingly called petite negative. The primary action of acriflavine, namely the inhibition of the synthesis of the respiratory system, was shown to occur not only in petite positive yeasts, but also in petite negative ones. Some implications of the results are discussed. 相似文献
4.
Seven regions of the subunit of human insulin receptor (HIR) were synthesized and examined for their ability to bind radioiodinated insulin. A peptide representing one of these regions (namely, residues 655–670) exhibited a specific binding activity for insulin. In quantitative radiometric titrations, the binding curves of125I-labeled insulin to adsorbents of peptide 655–670 and of purified placental membrane were similar or superimposable. The binding of radioiodinated insulin to peptide or to membrane adsorbents was completely inhibited by unlabeled insulin, and the inhibition curves indicated that the peptide and the membrane on the adsorbents had similar affinities. Synthetic peptides that were shorter (peptide 661–670) or longer (peptide 651–670) than the region 655–670 exhibited lower insulin-binding activity. It was concluded that an insulin-binding region in the HIR subunit resides within residues 655–670. The results do not rule out the possibility that other regions of the subunit may also participate in binding of HIR to insulin, with the region described here forming a face within a larger binding site. 相似文献
5.
Short peptides connecting-helices and-strands have been analyzed in 240 proteins refined at resolutions of 0.25 nm or better. Connecting peptides of lengths between one and five residues have been classified as part of supersecondary motifs of four types:, , , and. Careful consideration has been given to the definition of secondary structures on the basis of hydrogen bonds and main-chain conformational angles. Using five classes of residue conformation—a, b, e, l, t—in the nonregular structure regions of, space, 34 classes of supersecondary motifs occurring at least five times have been identified. Among these 34 classes, 11 classes that occur more than 25 times are commonly occurring supersecondary structure motifs. The patterns and conformations of the 11 commonly occurring supersecondary structure motifs have been characterized, demonstrating that patterns and conformations adopted by supersecondary structure motifs are limited. The results have relevance to structure prediction, comparative modeling, and protein folding. 相似文献
6.
Characterization of chromosomes with hybrid genes for Hb Lepore-Washington,Hb Lepore-Baltimore,Hb P-Nilotic,and Hb Kenya 总被引:1,自引:0,他引:1
K. D. Lanclos J. Patterson G. D. Efremov S. C. Wong A. Villegas P. J. Ojwang J. B. Wilson F. Kutlar T. H. J. Huisman 《Human genetics》1987,77(1):40-45
Summary This study concerns the characterization of chromosomes with hybrid genes for Hb Lepore-Washington (44 chromosomes), for Hb Lepore-Baltimore (5 chromosomes), for Hb P-Nilotic (8 chromosomes), and for Hb Kenya (7 chromosomes) by determining a relatively large number of restriction enzyme polymorphism. Two, and possibly three, different Hb Lepore-Washington chromosomes were identified by specific haplotypes, while the haplotype of the Hb Lepore-Baltimore chromosome had its own characteristic pattern. A likely conclusion is that the crossovers leading to the formation of these chromosomes have occurred as independent events within the populations. Chromosomes with the -Lepore-Washington hybrid gene maintained specific characteristies (such as increased Hb F levels in heterozygotes, and high or low G values in this Hb F) which have been observed in normal individuals with chromosomes having comparable haplotypes. Only one haplotype was observed for each of the chromosomes carrying either the -P-Nilotic hybrid gene or the A hybrid gene of Hb Kenya. 相似文献
7.
Stefan Andersson 《Plant Systematics and Evolution》1989,168(3-4):227-236
The extent of self-fertility was examined in 16 populations ofCrepis tectorum. A hypothesis that a weedy habit favours autogamy was only partly supported. Low levels of self-fertility characterized non-weedy populations from calcareous grasslands (alvars) on the Baltic island in Öland. By contrast, plants in nearly all weed populations studied were more or less self-fertile. However, the trend towards autogamy may have occurred independently of the trend towards a weedy habit, as shown by moderately to high levels of self-fertility in alvar populations from two other Baltic islands. In the weed group, there was a tendency for plants from two field populations to be more autogamous than plants from more ruderal habitats. There was an association between self-fertility and small, inconspicious heads in the alvar group but the association was weaker when weed populations were also considered. The relatively wide heads characterizing the ruderal weed populations may, at least partly, be an indirect effect of increases in overall plant size and/or in the size of the fruit associated with each flower. 相似文献
8.
The percentages of the -chain variant Hb G-Philadelphia (Hb G) or 2 68 AsnLys2 were evaluated in 84 adult and 18 newborn heterozygotes. These included members of three families who were studied in more detail by nucleic acid hybridization techniques. The adult heterozygotes fell in two categories, one with a higher proportion of Hb G [46.5±1.0% (SD), N=21] and another with lower values (33.9±3.4%, N=63). Among the newborn heterozygotes, two babies fell in the category with the higher proportion of Hb G while 16 babies gave values between 25 and 34%. Studies of -chain gene organization on the parents of one neonate with a Hb G level of 27% at birth and 37% at 8 months excluded the presence of chromosomes with triplicated -chain genes which could lead to the 0G/ genotype. Rather, these studies on five Hb G heterozygotes from three families confirmed the linkage between Hb G and a specific type of -thalassemia-2 associated with the presence of a 16-kbp Bgl II fragment which most probably carries the G locus since it has been found in 19 Hb G heterozygotes studied to date. The presence of an -thal-2 heterozygosity and three -chain genes (0G/) was confirmed among Hb G heterozygotes with lower proportions of this variant. It is likely that the even lower values found in some newborn could arise through defective assembly of G- dimers. The presence of an -thal-2 homozygosity and two active -chain genes, one on each chromosome (0G/0), was confirmed among heterozygotes with the higher proportion of Hb G. One of each of these categories was present in each of the three families investigated. This type of variability in the number of active -chain genes due to a heterozygosity or a homozygosity for -thalassemia-2 explains the trimodality of Hb S percentages among heterozygotes and the atypical hematological or biosynthetic features among patients with -thalassemia and sickle-cell syndromes.This research was supported by USPHS Research Grants HLB-05168 and HLB-15158 and by designated research funds of the Veterans Administration. This is Contribution No. 0693 of the Department of Cell and Molecular Biology, Medical College of Georgia, Augusta. 相似文献
9.
Horst Kessler Beate Diefenbach Dirk Finsinger Armin Geyer Marion Gurrath Simon L. Goodman Günter Hölzemann Roland Haubner Alfred Jonczyk Gerhard Müller Erich Graf von Roedern Jochen Wermuth 《Letters in Peptide Science》1995,2(3-4):155-160
Summary Integrins play a major role in cell-cell and cell-matrix interactions. The majority of the different types of integrins recognize the tripeptide sequence arginine-glycine-aspartic acid (RGD). To explore the spatial requirements of the pharmacophore for receptor selectivity and high activity, a new procedure, spatial screening, was used. The procedure is based on the experience that the conformation of small cyclic peptides is mainly determined by the chirality of the amino acids (and glycine or proline). For example, cyclic pentapeptides with one d and four l amino acids prefer a II'/ conformation. The sequence RGDFV was shifted around this spatial II'/ template by synthesis of five peptides in which one of the amino acids was used in d-configuration. It turned out that cyclo(-RGDfV-) is a selective inhibitor for the v3 integrin, which is strongly expressed in cancer cells. Systematic variations with different turn mimetics, retro-inverso structures, modified peptide bonds and sugar amino acids are discussed. 相似文献
10.
Andrea Calderan Paolo Ruzza Biancamaria Ancona Lorenzo Cima Pietro Giusti Gianfranco Borin 《Letters in Peptide Science》1998,5(2-3):71-73
Dermorphin structure–activity relationships toward and opioid receptors were investigated using a series of synthetic peptides, in which the aromatic residues at positions 1 or/and 3 of the N-terminal tetrapeptide analogue H-Tyr-d-Arg-Phe--Ala-NH2 were replaced by unnatural or constrained amino acids. 相似文献
11.
FLRFa, FIRFa, YMRFa, and some other peptides are found to slow down the kinetics of desensitization of proton-gated currents in rat trigeminal neurons but not in HEK-293T cell culture transfected with ASIC. The observed heterogeneity of peptide action in sensory neurons leads to a suggestion that FMRFa-related peptides may selectively affect transient current components of ASIC and DRASIC/DRASIC-MDEG2 heteromers. 相似文献
12.
13.
The kinetics of CNProto- and CNDeutero-hemin binding to apohemoglobin A2 was investigated in a stopped-flow device in 0.05 M potassium phosphate buffer, pH 7, at 10°C. The overall kinetic profile exhibited multiple phases: Phases I–IV corresponding with heme insertion (8.5–13 × 107 M–1 s–1), local structural rearrangement (0.21–0.23 s–1), global structural event (0.071–0.098 s–1), and formation of the Fe–His bond (0.009–0.012 s–1), respectively. Kinetic differences observed between apohemoglobin A2 and apohemoglobin A (previously studied) prompted an analysis of the structures of and chains through molecular modeling. This revealed a structural repositioning of the residues not only at, but also distant from the site of the amino acid substitutions, specifically those involved in the heme contact and subunit interface. A significant global change was observed in the structure of the exon-coded 3 region and provided additional evidence for the designation of this as the subunit assembly domain. 相似文献
14.
J. Visscher C. G. Bakker Alan W. Schwartz 《Origins of life and evolution of the biosphere》1990,20(5):369-375
A pyrophosphate-linked polynucleotide analog based on thymidine 3,5 bis-phosphate (pTp) catalyzes the oligomerization of activated dimers of pdAp in the presence of MgCl2. Although no catalysis of the oligomerization of the activated monomer (ImpdAplm) was observed in the presence of MgCl2, there was a significant stimulation of oligomerization by the template in the presence of MnCl2. 相似文献
15.
Heme metabolism in promastigotes of Leishmania donovani 总被引:3,自引:0,他引:3
Srivastava Pratima Sharma Guru Dutt Kamboj Kamal K. Rastogi Anil K. Pandey Vikas C. 《Molecular and cellular biochemistry》1997,171(1-2):65-68
Promastigotes of Leishmania donovani (Dd-8 strain) showed presence of important key enzymes of heme synthesizing (d-aminolevulinic acid synthase and ferrochelatase) and degrading (heme oxygenase and biliverdin reductase) systems, classical leishmanicidal drugs viz allopurinol, amphotericin B, pentamidine and CDRI compound 93/202 inhibited the heme oxygenase activity of the parasite, whereas, -aminolevulinic acid synthase activity practically remained unaffected. The Km, Vmax ad pH values of heme oxygenase of promastigotes were found to be 1666 M hemin, 625 nmol of bilirubin formed h-1 mg protein-1 and 7.5 respectively. The findings suggest the presence and importance of heme metabolism in the de novo synthesis of different hemoproteins of the Leishmania parasite as well as the detoxification and its defence against biological insults. 相似文献
16.
Pedro Garcia Anne-Marie Gasc Xenophon Kyriakidis Daniel Baty Michel Sicard 《Molecular & general genetics : MGG》1988,214(3):509-513
Summary In pneumococcal transformation a particular point mutation belonging to the amiA locus is able markedly to enhance recombination frequency when crossed with any other markers of this gene. This results from a polarized conversion of the mutation towards the wild-type sequence. In this report, by site-directed oligonucleotide mutagenesis, we have generated a series of mutants showing various degrees of conversion. We have found that the substitution 5-ATTCAT5-ATTAAT is a sufficient signal for localized conversion. Changing individual bases within this sequence results in decreased conversion frequencies to levels that depend on the mutation, suggesting that there is a family to related sequences which may act as a substrate for a conversion system. Moreover, the length over which this conversion occurs has been estimated to be 12 base pairs on the average. 相似文献
17.
Stephen A. Kuby Minoru Hamada Mark S. Johnson Geoffrey A. Russell Michael Manship Richard H. Palmieri Gerald Fleming David S. Bredt Albert S. Mildvan 《Journal of Protein Chemistry》1989,8(4):549-562
Two peptide fragments, derived from the head and tail of rabbit muscle myokinase, were found to possess remarkable and specific ligand-binding properties (Hamadaet al., 1979).By initiating systematic syntheses and measurements of equilibrium substrate-binding properties of these two sets of peptides, or portions thereof, which encompass the binding sites for (a) the magnesium complexes of the nucleotide substrates (MgATP2– and MgADP–) and (b) the uncomplexed nucleotide substrates (ADP3– and AMP2–) of rabbit muscle myokinase, some of the requirements for binding of the substrates to ATP-AMP transphosphorylase are being deduced and chemically outlined. One requirement for tight nucleotide binding appears to be a minimum peptide length of 15–25 residues. In addition, Lys-172 and/or Lys-194 may be involved in the binding of AMP.The syntheses are described as a set of peptides corresponding to residues 31–45, 20–45, 5–45, and 1–45, and a set of peptides corresponding to residues 178–192, 178–194, and 172–194 of rabbit muscle adenylate kinase. The ligand-binding properties of the first set of synthetic peptides to the fluorescent ligands: MgATP/ATP and MgADP/ADP are quantitatively presented in terms of their intrinsic dissociation constants (Kd) and values ofN (maximal number of moles bound per mole of peptide); and compared with the peptide fragment MT-I (1–44) obtained from rabbit muscle myokinase (Kubyet al., 1984) and with the native enzyme (Hamadaet al., 1979). In addition, the values ofN andKd are given for the second set of synthetic peptides to the fluorescent ligands AMP and ADP as well as for the peptide fragments MT-XII(172–194) and CB-VI(126–194) (Kuby et al., 1984) and, in turn, compared with the native enzyme.A few miscellaneous dissociation constants which had been derived kinetically are also given for comparison (e.g., theK
i for AMP and the value of
obtained for the native enzyme) (Hamada and Kuby, 1978), and theK'd measured for Cr3+ and the synthetic peptide I1–45 (Fryet al., 1985b).Paper XVII of this series is Kubyet al. (1983). 相似文献
18.
I. Tsekos 《Protoplasma》1996,193(1-4):10-32
Summary The relationship between the supramolecular organization of red algal cell membranes and the biosynthesis and secretion of the cell wall skeletal and matrix polysaccharides is reviewed. Freeze-fracture studies have revealed that organized macromolecular structures — linear terminal complexes and tetrads — are present on the plasma membrane and on membranes of the endomembrane system. The linear terminal complexes seem to be involved in the biosynthesis, assembly, and orientation of the cellulose microfibrils and the tetrads in the synthesis of the matrix polysaccharides. It is shown how the research on the supramolecular organization of cell membranes has increased the knowledge on the biosynthesis and secretion of the extracellular crystalline and non-crystalline polysaccharides in red algae. In this review, the progress to date is discussed.Dedicated to Prof. Dr. Dr. h.c. Eberhard Schnepf on the occasion of his retirement 相似文献
19.
Chen Jianwu Sun Fan Tang Lixia Tang Mujin Shi Yongxia Yu Jianxiu Pang Yi 《World journal of microbiology & biotechnology》2003,19(9):883-889
Vip3A is an 89-kDa protein secreted by Bacillus thuringiensis during vegetative growth. The 3.5 kb full-length vip184 gene was cloned from a wild-type isolate of B. thuringiensis, and the vip184S gene was constructed by deletion of the putative signal peptide encoding sequence. Both genes were expressed in the acrystalliferous strain Cry–B of B. thuringiensis. Vip184 protein was observed mainly in the centrifuged pellets of B. thuringiensis Cry–B(pHPT3), which contains the vip184 gene, and was less abundant in the concentrated supernatant. However, Vip184S proteins were not detected in the concentrated supernatant, but only in the pellets of Cry–B(pHPT3S), which contains vip184S gene. This indicated that Vip184S proteins were not secreted into the culture medium and that the putative signal peptides were essential for the secretion of Vip184. The toxicity of Cry–B(pHPT3) and Cry–B(pHPT3S) were demonstrated against the neonate larvae of Spodoptera exigua and S. litura. Pellets and concentrated supernatant of Cry–B(pHPT3) showed high activity against S. exigua and S. litura, but the Cry–B(pHPT3S) strain was not toxic to either because of the deletion of N-terminal putative signal peptides. Therefore, this may suggest that the putative signal peptides are required for lethality. 相似文献
20.
Summary Segregation analysis was performed on 211 nuclear families belonging to 166 pedigrees of hereditary retinoblastoma found in a number of series which have been gathered from the literature. Bilaterally affected carriers appear homogeneous. The segregation ratio in their offspring is 0.49, and the proportion of bilateral cases among affected offspring is 0.87. Both unilaterally affected and unaffected carriers appear heterogeneous. The very low segregation ratio (0.08) in the offspring of unilateral carriers who are not detected through an affected child, suggests the possiblity of two types of carriers, high and low transmitters. The proportions of low transmitters was estimated as 0.14 among all familial unilateral carriers and as 0.45 among all detected unaffected carriers. Unilateral and unaffected high transmitters give a significantly lower segregation ratio than bilaterally affected carriers.On the one hand, the existence of these two different types of carriers provides arguments in support of the hypothesis of delayed mutation. On the other hand, the differences in penetrance among high transmitters, according to their phenotype, supports the hypothesis of host resistance. Under the two-mutation hypothesis, the possibility that the mutation rate is variable among individuals and partly genetically determined, is suggested. 相似文献