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1.
Purification and characterization of a lysophospholipase from human amnionic membranes 总被引:1,自引:0,他引:1
We have identified the presence of a lysophospholipase in human placental tissues and have purified this enzyme from the amnion. The specific activity was highest in the amnion and decreased across adjacent tissues. The purification involved the use of DEAE-Sephadex, phenyl-Sepharose, hydroxylapatite, and sulfylpropyl Sephadex chromatography. The activity of the purified enzyme toward palmitoyl lysophosphatidylcholine is 2.5 mumol min-1 mg-1 and the pH optimum is 7.0. The enzyme is not inhibited by EDTA and does not appear to have a metal ion requirement. The enzyme may be of membrane origin; the purified enzyme requires the presence of detergent during storage. The effects of substrate composition and physical state on enzymatic activity were explored. The enzyme was not active toward mono-, di-, or triglycerides, nor toward diacyl phospholipid. The enzyme was active toward myristoyl and palmitoyl lysophosphatidylcholine at concentrations where these substrates spontaneously form micelles or where Triton X-100 was used to induce co-micellization of the substrate at low concentrations with detergent. A role for this enzyme in processing the lysophospholipid product of phospholipase A action must be considered in evaluating arachidonic acid production in human fetal membranes and placental tissue, particularly during the initiation of labor. 相似文献
2.
The effects of pTR2030 on the replication of four small isometric bacteriophages were examined in Streptococcus cremoris R1. Three lytic phages (652, 720, and 751), which were isolated independently over a 29-year period, were unable to form plaques on a pTR2030 transconjugant of S. cremoris R1. The fourth phage evaluated, phage r(1)t, was a temperate phage induced from S. cremoris R1 by treatment with mitomycin C. A prophage-cured derivative of S. cremoris R1, designated R1Cs, was isolated and served as a lytic indicator for phage r(1)t. Strain R1Cs and a derivative of this strain that was relysogenized with r(1)t, designated R1Cs(r(1)t), were used as conjugal recipients for transfer of the phage resistance plasmid pTR2030. pTR2030 transconjugants of strains R1Cs and R1Cs(r(1)t) were evaluated for sensitivity to r(1)t phage and induction of r(1)t prophage, respectively. The temperate phage r(1)t adsorbed eficiently but did not form plaques on the prophage-cured, pTR2030 transconjugant strain T-R1Cs. However, in the r(1)t lysogen [T-R1Cs(r(1)t)], pTR2030 did not inhibit prophage induction with mitomycin C, cell lysis, or production of infective r(1)t phage particles. The data demonstrated that pTR2030-induced resistance inhibited lytic infection by r(1)t phage from without but did not retard lytic development after prophage induction within the cell. It was suggested that pTR2030-encoded phage resistance to small isometric phages may, therefore, act at the cell surface or membrane to prevent phage DNA passage into the host cell or inhibit early events required for lytic replication of externally infecting phage. 相似文献
3.
Baulcombe DC Huttly AK Martienssen RA Barker RF Jarvis MG 《Molecular & general genetics : MGG》1987,209(1):33-40
Summary A genomic clone of a wheat -amylase gene (Amy3/33) was identified, on the basis of hybridisation properties, as different from -Amy1 and -Amy2 genes which had been characterised previously. The nucleotide sequence revealed that this gene has the normal sequence motifs of an active gene and an open reading frame interrupted by two introns. The protein sequence encoded by this open reading frame is recognisably similar to that of -amylase from the -Amy1 and -Amy2 genes and there is high sequence homology in all three proteins at the putative active sites and Ca++ binding region. In addition, the introns are at positions equivalent to the position of introns in the -Amy1 and -Amy2 genes. However, the sequence was less similar to -Amy1 and -Amy2 than these are to each other. Southern blot analysis showed that the Amy3/33 DNA is one of a small multigene family carried on a different chromosome (group 5) from either the -Amy1 or -Amy2 genes. A further difference from the -Amy1 and -Amy2 genes was the pattern of expression. Amy3/33 was expressed only in immature grains and, unlike the -Amy1 and -Amy2 genes, not at all in germinating aleurones. These data suggested therefore that this gene represents a third type of -amylase gene, not described before, which shares a common evolutionary ancestor with the -Amy1 and -Amy2 genes. 相似文献
4.
Differences in the chemical and catalytic characteristics of two crystallographically ''identical'' enzyme catalytic sites. Characterization of actinidin and papain by a combination of pH-dependent substrate catalysis kinetics and reactivity probe studies targeted on the catalytic-site thiol group and its immediate microenvironment. 总被引:5,自引:5,他引:0 下载免费PDF全文
E Salih J P Malthouse D Kowlessur M Jarvis M O''Driscoll K Brocklehurst 《The Biochemical journal》1987,247(1):181-193
The characteristics of actinidin (EC 3.4.22.14) and papain (EC 3.4.22.2), two cysteine proteinases whose catalytic-site regions appear to superimpose to a degree that approaches atomic co-ordinate accuracy of both crystal structures, were evaluated by determining (a) the pH-dependence in acid media of the acylation process of the catalytic act (k+2/Ks) using N alpha-benzoyl-L-arginine p-nitroanilide (L-Bz-Arg-Nan) as substrate and (b) the sensitivity of the reactivity of the catalytic-site thiol group and its pH-dependence to structural change in small, thiol-specific, two-protonic-state reactivity probes (2,2'-dipyridyl disulphide and methyl 2-pyridyl disulphide) where enzyme-probe contacts should be restricted to areas close to the catalytic site. Distortion of the catalytic sites of the two enzymes at pH less than 4 was evaluated over time-scales appropriate for both stopped-flow reactivity probe kinetics (less than or equal to 1-2 s) and steady-state substrate catalysis kinetics (3-5 min) by using the 2,2'-dipyridyl disulphide monocation as a titrant for non-distorted catalytic sites. This permitted a lower pH limit to be defined for valid kinetic analysis of both types. The behaviour of the enzymes at pH less than 4 requires a kinetic model in which the apparently biomolecular reaction of enzyme with probe reagent is separated from the process leading to loss of conformational integrity by a potentially reversible step. The acylation of actinidin with L-Bz-Arg-Nan in acidic media occurs in two protonic states, one produced by raising the pH across pKa less than 4 which probably characterizes the formation of -S-/-ImH+ ion pair (pKa approx. 3) and the other, of higher reactivity, produced by raising the pH across pKa 5.5, which may characterize rearrangement of catalytic-site geometry. The pH-dependence of the acylation of papain by L-Bz-Arg-Nan is quite different and is not influenced by protonic dissociation with pKa values in the range 5-6. The earlier conclusion that the acylation of papain depends on two protonic dissociations each with pKa approx. 4 was confirmed. This argument is now more firmly based because titration with 2,2'-dipyridyl disulphide permits the loss of conformational integrity to be taken into account in the analysis of the kinetic data at very low pH. Methyl 2-pyridyl disulphide was synthesized by reaction of pyridine-2-thione with methyl methanethiolsulphonate and its pKa at I = 0.1 was determined by spectral analysis at 307 nm to be 2.8.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
5.
Grain yields were measured over 2 seasons from a range of field crops following liming and deep ripping an acid and compacted soil in north-eastern Victoria. Lime (2.5 t ha–1) substantially reduced the level of exchangeable Al and exchangeable Mn whilst raising soil pH by about 1.0 unit. The crops grown were 7 cultivars of wheat and one cultivar each of triticale, oats, barley, rapeseed, safflower, field pea, chick pea and lupins. With the exception of lupin, liming the soil increased (p=0.05) the grain yield of all crops and cultivars. With the wheat cultivars there were 2 distinct groups with different tolerance to soil acidity. Wheat, oats, triticale and lupins had higher absolute yields than the other crops. Safflower and chick pea had very low yields without soil amendment. The magnitude of the lime response did not differ between the wheat cultivars (17%) or between any of the crop species (range 9–29%). Deep ripping the soil to break a hard compacted layer resulted in more yield for all the cereals and safflower. The results demonstrate the importance of using crops with tolerance to acid soil conditions as well as gains that can be obtained with ameliorating identifiable soil problems. 相似文献
6.
M.C. Jarvis 《Letters in applied microbiology》1988,7(6):157-159
Mycelial cell walls from the potato blight fungus ( Phytophthora infestans ) (Mont.) de Bary were examined in the solid state by 13 C nuclear magnetic resonance spectroscopy with cross-polarization and magic-angle spinning. The spectrum was free from interference by spinning sidebands. The main component of the cell walls had the spectral properties of a β -(1,3')-glucan. Protein appeared to be present also. The presence of β -(1,6')-linked glucose residues, cellulose or chitin was not ruled out, but there was no evidence for these as major components of the cell walls. 相似文献
7.
Peter M. Narins O. J. Reichman Jennifer U. M. Jarvis Edwin R. Lewis 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1992,170(1):13-21
Summary Both seismic and auditory signals were tested for their propagation characteristics in a field study of the Cape mole-rat (Georychus capensis), a subterranean rodent in the family Bathyergidae. This solitary animal is entirely fossorial and apparently communicates with its conspecifics by alternately drumming its hind legs on the burrow floor. Signal production in this species is sexually dimorphic, and mate attraction is likely mediated primarily by seismic signalling between individuals in neighboring burrows. Measurements within, and at various distances away from, natural burrows suggest that seismic signals propagate at least an order of magnitude better than auditory signals. Moreover, using a mechanical thumper which could be triggered from a tape recording of the mole-rat's seismic signals, we established that the vertically-polarized surface wave (Rayleigh wave) propagates with less attenuation than either of the two horizontally-polarized waves. Thus, we tentatively hypothesize that Rayleigh waves subserve intraspecific communication in this species.Abbreviations
PPM
pulses per min
-
SB
simulated burrow
-
SD
standard deviation
-
SPL
sound pressure level 相似文献
8.
The common mole-rat, Cryptomys h. hottentotus , is a social subterranean rodent occurring in colonies in which one female and one to three males are involved in reproduction and the remaining colony members are non-reproductive. Within each sex the reproductive animals are usually the largest and most dominant animals.
The dominance hierarchy amongst a field-captured colony was linear ( h = 0.95, calculated from Landau's linearity index) soon after capture. The non-reproductive females were ranked low in the dominance hierarchy; many were subordinate to non-reproductive males. The order of capture of mole-rats was not related to the position in the dominance hierarchy. The hierarchy became non-linear ( h = 0.56) after six months in captivity during which two juvenile animals became adult. The breakdown in the hierarchy may result from the lack of opportunity in captivity for animals to disperse and establish satellite colonies, or from colony members becoming co-dominant in the hierarchy as a result of a rise in rank by young animals.
Dominant mole-rats are involved in a greater proportion of interactive behaviours than subordinates. Popularity studies show that females tend to be more popular animals than males. The largest reproductive male was the least popular animal in the first study, whereas a beta male was the least popular animal in the second study period. The reproductive female was the most popular in both periods. 相似文献
The dominance hierarchy amongst a field-captured colony was linear ( h = 0.95, calculated from Landau's linearity index) soon after capture. The non-reproductive females were ranked low in the dominance hierarchy; many were subordinate to non-reproductive males. The order of capture of mole-rats was not related to the position in the dominance hierarchy. The hierarchy became non-linear ( h = 0.56) after six months in captivity during which two juvenile animals became adult. The breakdown in the hierarchy may result from the lack of opportunity in captivity for animals to disperse and establish satellite colonies, or from colony members becoming co-dominant in the hierarchy as a result of a rise in rank by young animals.
Dominant mole-rats are involved in a greater proportion of interactive behaviours than subordinates. Popularity studies show that females tend to be more popular animals than males. The largest reproductive male was the least popular animal in the first study, whereas a beta male was the least popular animal in the second study period. The reproductive female was the most popular in both periods. 相似文献
9.
E. D. Jarvis R. L. Widom G. LaFauci Y. Setoguchi I. R. Richter R. Rudner 《Genetics》1988,120(3):625-635
Integrative mapping with vectors containing ribosomal DNA sequences were used to complete the mapping of the 10 rRNA gene sets in the endospore forming bacterium Bacillus subtilis. Southern hybridizations allowed the assignment of nine operons to distinct BclI restriction fragments and their genetic locus identified by transductional crosses. Nine of the ten rRNA gene sets are located between 0 and 70 degrees on the genomic map. In the region surrounding cysA14, two sets of closely spaced tandem clusters are present. The first (rrnJ and rrnW) is located between purA16 and cysA14 closely linked to the latter; the second (rrnI, rrnH and rrnG) previously mapped within this area is located between attSPO2 and glpT6. The operons at or near the origin of replication (rrnO,rrnA and rrnJ,rrnW) represent "hot spots" of plasmid insertion. 相似文献
10.
Many laboratory strains of Bacillus subtilis contain 9 rather than 10 rRNA operons due to deletions occurring within the rrnJ-rrnW or rrnI-rrnH-rrnG gene cluster. These operons are members of two sets of closely spaced clusters located in the cysA-aroI region. Analysis of rescued DNA from integrants with insertions into rrnG and rrnH indicated that these tandemly arranged operons allowed frequent deletions of an rrn operon equivalent. These events may arise spontaneously by intrachromosomal recombination or by simultaneous double crossovers with a multimeric integrative plasmid. 相似文献