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1.
This review summarizes and analyzes data on structural and functional peculiarities of - and -subunits of heterotrimeric G proteins and their participation in -dimer in processes of hormonal signal transduction to the cell. The molecular mechanisms are discussed, which are responsible for formation of the -dimer complex and its interaction with the wide spectrum of signal and structural proteins, such as G protein -subunit, serpentine type receptors, kinases of G protein-coupled receptors, adenylyl cyclase, phospholipase C2, phosphatidyl-inositol-3-kinase, Raf-kinase, potassium and calcium channels, microtubular and RGS-proteins. The characteristics of - and -subunits are given for living organisms of different phylogenetic levels (lower eukaryotes, fungi, plants, vertebrates, and invertebrates). It is concluded that the -dimer is the most important regulatory link in various signal systems to carry out switch from one signal cascade to another and thereby to determine the final cellular response to the signal impulse.  相似文献   

2.
The relative roles of the two structural aspects of nonenzymic glycation sites of hemoglobin A, namely the ease with which the amino groups could form the aldimine adducts and the propensity of the microenvironments of the respective aldimines to facilitate the Amadori rearrangement, in dictating the site selectivity of nonenzymic glycation with aldotriose has been investigated. The chemical reactivity of the amino groups of hemoglobin A forin vitro reductive glycation with aldotriose is distinct from that in the nonreductive mode. The reactivity of amino groups of hemoglobin A toward reductive glycation (i.e., propensity for aldimine formation) decreases in the order Val-1(), Val-1(), Lys-66(), Lys-61(), and Lys-16(). The overall reactivity of hemoglobin A toward nonreductive glycation decreased in the order Lys-16(), Val-1(), Lys-66(), Lys-82(), Lys-61(), and Val-1(). Since the aldimine is the common intermediate for both the reductive and nonreductive modification, the differential selectivity of protein for the two modes of glycation is clearly a reflection of the propensity of the microenvironments of nonenzymic glycation sites to facilitate the isomerization reaction (i.e., Amadori rearrangement). A semiquantitative estimate of this propensity of the microenvironment of the nonenzymic glycation sites has been obtained by comparing the nonreductive (nonenzymic) and reductive modification at individual glycation sites. The microenvironment of Lys-16() is very efficient in facilitating the rearrangement and the relative efficiency decreases in the order Lys-16(), Lys-82(), Lys-66(), Lys-61(), Val-1(), and Val-1(). The propensity of the microenvironment of Lys-16() to facilitate the Amadori rearrangement of the aldimine is about three orders of magnitude higher than that of Val-1() and is about 50 times higher than that of Val-1(). The extent of nonenzymic glycation at the individual sites is modulated by various factors, such as thepH, concentration of aldotriose, and the concentration of the protein. The nucleophiles—such as tris, glycine ethyl ester, and amino guanidine—inhibit the glycation by trapping the aldotriose. The nonenzymic glycation inhibitory power of nucleophile is directly related to its propensity to form aldimine. Thus, the extent of inhibition of nonenzymic glycation at a given site by a nucleophile directly reflects the relative role ofpK a of the site in dictating the glycation at that site. The nonenzymic glycation of an amino group of a protein is an additive/synergestic consequence of the propensity of the site to form aldimine adducts on one hand, and the propensity of its microenvironment to facilitate the isomerization of the aldimines to ketoamines on the other. The isomerization potential of microenvironment plays the dominant role in dictating the site specificity of the nonenzymic glycation of proteins.  相似文献   

3.
Unravelling the factors that contribute to the formation and the stability of -sheet structure in peptides is a subject of great current interest. A -hairpin, the smallest -sheet motif, consists of two antiparallel hydrogen-bonded -strands linked by a loop region. We have performed a statistical analysis on protein -hairpins showing that the most abundant types of -hairpins, 2:2, 3:5 and 4:4, have characteristic patterns of 13C and 13C conformational shifts, as expected on the basis of their and angles. This fact strongly supports the potential value of 13C and 13C conformational shifts as a means to identify -hairpin motifs in peptides. Their usefulness was confirmed by analysing the patterns of 13C and 13C conformational shifts in 13 short peptides, 10–15 residues long, that adopt -hairpin structures in aqueous solution. Furthermore, we have investigated their potential as a method to quantify -hairpin populations in peptides.  相似文献   

4.
Zusammenfassung Junge Halsbandschnäpper wurden handaufgezogen, flogen im Flugkäfig aus und wurden dort selbständig. Darauf wurden sie 90 km nach Süden verfrachtet und in einem von dieser Art unbewohnten Gebiet freigelassen. Im nächsten Frühjahr siedelten sich mindestens 9 dort an, was 19% Rückkehrern entspricht, wenn die Hälfte der Vögel waren. kehrten in geringerer Zahl zurück und wurden nicht restlos erfaßt.Eine weitere Gruppe wurde erst vor Ende der Jugendmauser verfrachtet. Auch davon kehrten 18-19% der zurück. Ein Zeitraum von rund 2 Wochen vor dem Wegzug reichte also zur Prägung auf ein Gebiet als Heimat aus.Von einer dritten Gruppe von insgesamt 68 Schnäppern (= ca. 34 ), die erst nach Ende der Jugendmauser zur Wegzugzeit aufgelassen wurde, konnte später keiner nachgewiesen werden, auch nicht am Aufzuchtsort. Letzteres könnte an der Ungunst der örtlichen Verhältnisse liegen.Mit Unterstützung der Deutschen Forschungsgemeinschaft.  相似文献   

5.
Summary Crude homogenate of thermophilic archaebacteriumSulfolobus solfataricus, possessing a -glycosidase, has been used to synthesize different alkyl -D-glycosides starting from phenyl -D-glucoside, phenyl -D-galactoside and lactose as carbohydrate donors. High product yield (95% with respect to the carbohydrate donor) of octyl -D-glucoside has been obtained in a two-phase system containing 5% of water. The enantioselection for the galactosyl transfer to the secondary hydroxyl group of propane-1,2-diol is higher than that found using -galactosidase fromE. coli.  相似文献   

6.
Summary We treated thirty seven patients suffering from a clinical stage I malignant melanoma by immunosurgery and observed them for a three-year period ending December 1975. Prior to surgery the tumor was submitted to the application of DNCB. Patients were divided between those who had or had not been subjected to biopsy before starting treatment. Accordingly, 23 patients treated by immunosurgery as the first treatment, were called the primitive treatment group and 14 patients subjected to a previous biopsy, the secondary treatment group. In the primitive treatment group, all 23 patients survived without any sign of recurrence. Relapse was observed in 4 out of 14 patients of the biopsy group. Extensive histological investigations have been made by light and electron microscopes. The tumors were infiltrated and disrupted by a large number of mononucleated cells. The majority of infiltrating cells were small lymphocytes. True lymphatic follicles have been observed in the skin around the melanoma.  相似文献   

7.
A novel syrup containing neofructo-oligosaccharides was produced from sucrose (Brix 70) by whole cells of Penicillium citrinum. The efficiency of fructo-oligosaccharides production was more than 55% and those of the main carbohydrate components, 1-kestose (Fruf 21Fruf 21 Glc), nystose (Fruf 21Fruf 21 Fruf 21 Glc) and neokestose (Fruf 26 Glc12 Fruf), were 22, 14 and 11%, respectively.  相似文献   

8.
A model of associative memory for time varying spatial patterns is proposed and simulated on a digital computer. This is a network composed of many neuron-like elements, and shows an ability for associative memory similar to that of the brain.Suppose a number of sequences of spatial patterns are presented to this network, for example, 12345, ABC, and so on. Then, these patterns are memorized in the network. After that, if any part of one of these sequences, say 23, is presented to the circuit, the rest of the sequence, 45, is recalled following to it. It resembles to such a situation — if we hear a part of a melody which we have memorized in the past, the rest of the melody is recalled even after it is stopped half-way. Although the recalled patterns are not always 100% correct, they are not completely destroyed even if the presented patterns are imperfect.  相似文献   

9.
The effect of inorganic phosphate, ADP, ATP, and their analogues on the rate of labeling of F1-ATPase by 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) and phenylglyoxal have been investigated. Analysis of the kinetic data indicate that the labeled functional groups of the essential tyrosine and arginine residues respectively are both located at the catalytic site of F1. The active phenolic group of tyrosine is located closer to the bound inorganic phosphate or the -phosphate group than the - and -phosphate groups of the bound ATP at the catalytic site, whereas the guanidinium group of arginine is located closer to the - and -phosphate groups of the bound ATP than to its -phosphate group or the bound inorganic phosphate. The kinetically deduced dissociation constants are 1.3 mM and 210 µM for the inorganic phosphate and ADP respectively bound to this catalytic site. Labeling the essential tyrosine residue by NDB-Cl has been found to facilitate subsequent labeling of the essential arginine residue by phenylglyoxal.Abbreviations NBD-Cl 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (this compound has been named 4-chloro-7-nitro-benzofurazan and abbreviated NBf-Cl elsewhere) - DTT dithiothreitol - EDTA ethylenediaminetetraacetic acid - Pi inorganic phosphate - PEP phosphoenolpyruvate - ADPCP ,-methylene-adenosine 5-triphosphate - AMPCP ,-methylene-adenosine 5-diphosphate - Hepes N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - Tris 2-amino-2(hydroxymethyl)-1,3-propanediol  相似文献   

10.
Ray  I.  Chauhan  A.  Wisniewski  H. M.  Wegiel  J.  Kim  K. S.  Chauhan  V. P. S. 《Neurochemical research》1998,23(10):1277-1282
Amyloid beta-protein (A), in its soluble form, is known to bind several circulatory proteins such as apolipoprotein (apo) E, apo J and transthyretin. However, the binding of A to intracellular proteins has not been studied. We have developed an overlay assay to study A binding to intracellular brain proteins. The supernatants from both rat and human brains were found to contain several proteins that bind to A 1–40 and A 1–42. No major difference was observed in the A binding-proteins from brain supernatants of patients with Alzheimer's disease and normal age-matched controls. Binding studies using shorter amyloid beta-peptides and competitive overlay assays showed that the binding site of A to brain proteins resides between 12–28 amino acid sequence of A. The presence of several intracellular A-binding (AB) proteins suggests that these proteins may either protect A from its fibrillization or alternatively promote A polymerization. Identification of these proteins and their binding affinities for A are needed to assess their potential role in the pathogenesis of Alzheimer's disease.  相似文献   

11.
Summary The histochemical activities of succinic dehydrogenase (SDH), myofibrillar Adenosine triphosphatase (ATPase) and alpha glycerophosphate dehydrogenase were studied in serial sections of rat vastus lateralis (red) (RVL), gastrocnemius and diaphragm muscles. Three main fibre-types were distinguished. The Type I fibres of RVL and gastrocnemius muscles fell into two distinct groups: one category-Type IA showed very low ATPase activity. The second category of Type IB fibres displayed moderate ATPase reaction. The Type IA fibres were divisible into two sub-groups when tested for SDH reaction. Type IA1 fibres possessed a homogenous distribution of diformazan·granules throughout the fibre: Type IA2 fibres displayed characteristic moth-eaten pattern of diformazan localization. The diaphragm muscle did not show either Type IB or Type IA2 varieties. The great majority of Type I fibres were sub-type IA1 in the three fast muscles studied. It is also demonstrated here that an inherent heterogeneity exists between Type I fibres of diaphragm and leg muscles in regard to -GPD localization. This histochemical data emphasizes the fact that subdivision of Type I striated muscle fibres of mammalian animals into two sub-types is only approximate and that a further subcategorization is possible.  相似文献   

12.
Summary Four types of striated muscle fibers with distinctive ultrastructure were defined in the Atlantic hagfish (Myxine glutinosa, L.): white, intermediate, and red fibers of m. parietalis, and red fibers of m. craniovelaris.White fibers are thick, contain very few mitochondria and fat vacuoles, and possess distinct and separate myofibrils with thin Z-disks and distinct M-lines. Intermediate fibers are thinner, possess largely similar myofibrils that often are even better separated due to a higher content of fat vacuoles and especially mitochondria and glycogen granules. Red fibers of m. parietalis contain large amounts of mitochondria, fat vacuoles, and glycogen granules. Their myofibrils possess M-lines, and although branching more, the myofibrils of red fibers conform with a Fibrillenstruktur pattern like those of white and intermediate fibers. Red fibers of m. craniovelaris are very thin, possess many smaller fat vacuoles, and large amounts of mitochondria and glycogen granules. The myofibrils are significantly thinner than in m. parietalis fibers, run as quite independent well separated units, possess thicker Z-disks, and lack M-lines. Large amounts of myosatellite cells are associated with these red fibers.Triads are located near A/I-junctions in all four fiber types and occur irregularly, the density of triads being different in the various fiber types.We are indebted to Dr. Finn Walvig, Biological Station, University of Oslo, Drøbak, for supply of hagfishes, and we also wish to thank Dr. Jan K. S. Jansen, Institute of Physiology, University of Oslo, for valuable suggestions during this study.  相似文献   

13.
Summary Two specific -N-acetylglucosaminyltransferases involved in the branching and elongation of mucin oligosaccharide chains, namely, a 1,6 N-acetylglucosaminylsaminyltransferase that transfers N-acetylglucosamine from UDP-N-acetylglucosamine to Gal3GalNAc-Mucin to yield Gal3(GlcNAc6)GalNAc-Mucin and a 3-N-acetylglucosaminyl transferase that transfers N-acetylglucosamine from UDP-N-acetylglucosamine to Gal3(GlcNAC6)GalNAc-mucin to yield GlcNAc3Gal3 (GlcNAc6)GalNAc-Mucin were purified from the microsomal fraction of swine trachea epithelium. The 1,6-N-acetylglucosaminyltransferase was purified about 21,800-fold by procedures which included affinity chromatography on DEAE columns containing bound asialo Cowper's gland mucin glycoprotein with Gal1,3GalNAc side chains. The apparent molecular weight estimated by gel filtration was found to be about 60 Kd. The purified enzyme showed a high specificity for Gal1,3GalNAc chains and the most active substrates were mucin glycoproteins containing these chains. The apparent Km of the 6-glucosaminyltrans-ferase for Cowper's gland mucin glycoprotein containing Gal1,3GalNAc chains was 0.53 µM; for UDP-N-acetylglucosamine, 12 µM; and for Gal 1,3GalNAc NO2ø, 4 mM. The activity of the 6-glucosaminyltransferase was dependent on the extent of glycosylation of the Gal3GalNAc chains in Cowper's gland mucin glycoprotein.The best substrate for the partially purified 3-Glucosaminyltransferase was Cowper's gland mucin glycoprotein containing Gal1,3(GlcNAc6)GalNAc side chains. This enzyme showed little or no activity with intact sialylated Cowper's gland mucin glycoprotein or derivatives of this glycoprotein containing GalNAc or Gal1,3GalNAc side chains.The radioactive oligosaccharides formed by these enzymes in large scale reaction mixtures were released from the mucin glycoproteins by treatment with alkaline borohydride, isolated by gel filtration on Bio-Gel P-6 and characterized by methylation analysis and sequential digestion with exoglycosidases. The oligosaccharide products formed by the 6- and 3-glucosaminyltransferases were shown to be Gal3(GlcNAC6) GalNAc and GlcNAc3 Gal3(GlcNAC6)GalNAc respectively.Taken collectively, these results demonstrate that swine trachea epithelium contains two specific N-acetylglucosaminyltransferases which catalyze the initial branching and elongation reactions involved in the synthesis of O-linked oligosaccharide chains in respiratory mucin glycoproteins. The first enzyme a 6-glucosaminyltransferase converts Gal3GalNAc chains in mucin glycoproteins to Gal3(GlcNAc6)GalNAc chains. This product is the substrate for a second 3-glucosaminyltransferase which converts the Gal3(GlcNAc6)GalNAc chains to GlcNAc3Gal(GlcNAc6)GalNAc chains in the glycoprotein. The 3-glucosaminyltransferase did not utilize Gal3GalNAc chains as a substrate and this results in an ordered sequence of addition of N-acetylglucosamine residues to growing oligosaccharide chains in tracheal mucin glycoproteins.Abbreviations NeuNAc N-acetylneuraminic acid - GalNAcol N-acetylgalactosaminitol - CGMG Cowper's gland mucin glycoprotein - GalNAc-CGMG Cowper's gland mucin glycoprotein containing GalNAc side chains O-glycosidically linked to serine or threonine - Gal3GalNAc-CGMC Cowper's gland mucin glycoprotein containing Gal3GalNAc side chains - MES 2-(N-morpholino) Ethane Sulfonic acid - PBS Phosphate Buffered Saline  相似文献   

14.
Gene targeting in embryonic stem (ES) cells via homologous recombination can occur at very low frequency. In order to enrich homologous recombinants before screening, a negative selection marker, such as thymidine kinase (TK) and diphtheria toxin A fragment (DTA), has been commonly used. In this study, we developed a negative selection marker using DTA gene with polyadenylation signal and it was designated DTApA. To determine the difference in targeting efficiency of the negative selections, we constructed three different targeting vectors for each negative selection (first, TK at the 3 end, second, TK at both the 5 and 3 ends <2 X TK>, and third, DTApA at the 5 end of the homologous sequences). Gene targeting experiments using these constructs clearly showed that negative selection using DTApA was more efficient than that using TK for homologous recombination and that negative selection using DTApA was as efficient as that using 2 X TK. Considering the fact that the use of DTApA is more convenient for construction of targeting vectors than that of 2 X TK, DTApA is an efficient negative selection marker.In addition, we examined long and accurate PCR (LAPCR) for screening gene targeted clones. The use of LAPCR with genomic DNAs from ES cell clones facilitated simple detection of homologous recombinants, suggesting that the screening with LAPCR is compatible with the use of longer homologous sequences of both arms in vector design. Our results indicate that the use of DTApA for negative selection together with the application of LAPCR for screening ensures efficient and timesaving screening for homologous recombinants.  相似文献   

15.
In addition to several cytochromes three iron sulfur proteins were detected in mixotrophically grown cells of Ectothiorhodospira shaposhnikovii, a member of the Chromatiaceae. They were identified as a bacterial ferredoxin and two high potential iron sulfur proteins (HIPIPs). The two HIPIPs were purified and characterized. They were named according to their differing retention times on a DEAE-cellulose column using a continuous NaCl gradient: early and late HIPIP. The HIPIPs contain 4 mol of non-heme iron and 4 mol of acid labile sulfur per mol protein. Under the conditions of purification the early HIPIP (E m, 7+270 mV) was present in a semi-reduced state. Using ion-exchange chromatography the early HIPIP could be split into a reduced green-brown (pI=3.7) and an oxidized red-brown (pI=3.9) fraction. The late HIPIP (pI=3.8) showed a midpoint potential of only+155 mV, the lowest redox potential of a HIPIP described so far.Non-common abbreviations HIPIP high potential iron sulfur protein - MOPS 3(N-morpholino)propane sulfonate - SDS sodium dodecylsulfate  相似文献   

16.
The primary structure of Rose-ringed Parakeet hemoglobin -chain was established, completing the analysis of this hemoglobin. Comparisons with other avian -chains show variations smaller than those for the corresponding -chains. There are 11 amino acid exchanges in relationship to the only other characterized psittaciform -chain, and a total of 35 positions are affected by differences among all avian -chains analyzed (versus 61 for the -chains). At three positions, the Psittacula -chain has residues unique to this species. Three 11 contacts are modified, by substitutions at positions 51, 116, and 125.  相似文献   

17.
Summary Activation of the -adrenergic receptors of the opercular epithelium ofFundulus heteroclitus stimulates Cl secretion, while activation of the -adrenergic receptors inhibits Cl secretion (Degnan and Zadunaisky, 1979). The possible involvement of adenosine 3, 5-monophosphate (cAMP) in these adrenergic responses was investigated. Isolated opercular epithelia incubated in Ringer, containing 10 mM theophylline, had cAMP levels ranging between 5.3 and 19.3 pmoles·mg protein–1 (mean=9.5±1.0 pmoles·mg protein–1). Activation of the -receptors by 10–5 M isoproterenol increased the mean cAMP level 430% (P<0.001). Blockage of the -receptors with propranolol greatly reduced the increase in cAMP in response to isoproterenol. Activation of the -receptors by 10–5 M arterenol stimulated the mean cAMP level 270% (P<0.01). However, when the -receptors were blocked with propranolol, arterenol had no effect on the cAMP level. The possible involvement of Ca++ in these adrenergic responses was investigated. Neither the stimulatory effect of isoproterenol, nor the inhibitory effect of arterenol on the Cl secretion were diminished in the absence of extracellular Ca++. The Ca++ ionophore, A23187, and the calmodulin inhibitor, trifluoperazine, had no effects on the Cl secretion. The Ca++-channel blocker, D600, had a significant inhibitory effect (P<0.005). Guanosine 3,5-monophosphate (cGMP) had no effect on the Cl secretion.The results indicate that -adrenergic stimulation of Cl secretion across the opercular epithelium is accompanied by an elevation in tissue cAMP levels. -adrenergic inhibition of Cl secretion does not involve changes in the tissue cAMP. Neither of these responses appear to require Ca++.  相似文献   

18.
In microbiology the terms viability and culturability are often equated. However, in recent years the apparently self-contradictory expression viable-but-nonculturable (VBNC) has been applied to cells with various and often poorly defined physiological attributes but which, nonetheless, could not be cultured by methods normally appropriate to the organism concerned. These attributes include apparent cell integrity, the possession of some form of measurable cellular activity and the apparent capacity to regain culturability. We review the evidence relating to putative VBNC cells and stress our view that most of the reports claiming a return to culturability have failed to exclude the regrowth of a limited number of cells which had never lost culturability. We argue that failure to differentiate clearly between use of the terms viability and culturability in an operational versus a conceptual sense is fuelling the current debate, and conclude with a number of proposals that are designed to help clarify the major issues involved. In particular, we suggest an alternative operational terminology that replaces VBNC with expressions that are internally consistent.  相似文献   

19.
Summary The winter wheat varieties Starke and Cappelle Desprez and the spring wheat Chinese Spring were analysed for structural chromosome rearrangements that resulted in the formation of multivalents in F1 hybrids. The analyses were carried out using hybrids involving euploids, monosomic and ditelosomic stocks, and double-monotelodisomic constructs. The study confirmed that Cappelle Desprez differs from Chinese Spring in a reciprocal translocation between chromosomes 5B and 7B (Riley et al. 1967); a translocation involving chromosomes 3B and 3D could not be verified. Furthermore, the analysis showed that Starke differs from Chinese Spring in a reciprocal translocation between chromosomes 7A and 7D. Both translocations have a coefficient of multivalent realisation of about 0.84. Further multivalents in euploid Starke, in euploid and some aneuploid stocks of Cappelle Desprez, and in euploid as well as various types of aneuploid hybrids between all three varieties could nearly all be explained hypothesizing that chromosome 2B of both Starke and Cappelle Desprez is a duplication-deficiency chromosome. In the hypothesis a part of the long arm of 2B is missing and replaced by a duplicated part of the long arm of chromosome 2D. The multivalents of this rearrangement showed an average coefficient of realisation of about 0.09.Sven Ellerström died in December 1985  相似文献   

20.
In Fuji, the production of ethylene was increased with the addition of AgNO3 and inhibited with the addition of 10 M aminoethoxyvinylglycine (AVG). The addition of 80 M AgNO3 to transformed explants of Fuji cultured on selection medium resulted in increased ethylene production (20 l l–1) at 3 weeks. Under examining the effect of AgNO3 in Fuji, the 40 M AgNO3 showed with higher 33.8% and 6.5% in the efficiency of regeneration and transformation. However, ethylene production in Gala explants treated with 10M AgNO3 (3 l l–1) decreased after 2 weeks compared with the control (5 l l–1). Although the regeneration efficiency of Gala with 10 M AgNO3 was higher (41.1%) than the control (20.1%), there was no significant difference in the transformation efficiency at the same concentration. Shoot regeneration of Fuji and Gala was completely inhibited with 10 M AVG. These results suggest that the addition of AgNO3 affects the efficiency of Agrobacterium-mediated gene transfer in Fuji.Eun Soo Seong, Ill Min Chung- These two Authors Contributed equally to this work  相似文献   

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