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1.
Menno P. Gerkema Floris van der Leest 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1991,168(5):591-597
Summary The timing mechanism underlying ultradian (2–3 h) activity patterns in the common vole, Microtus arvalis, was studied using behavioural deprivation experiments. These were aimed at distinguishing between a homeostatic control mechanism, in which the rhythmic behaviour itself is part of the causal loop, and a clock mechanism, independent of the behaviour.In 175 experiments, deprivation of food during 3 ultradian cycles in (subjective) daytime did not result in significant changes in the ultradian periodicity of attempts to obtain the food, compared with ad lib. access to food and water. A minor, but significant increase in ultradian activity time () occurred in the course of the deprivation, but this was compensated by a shorter ultradian rest (). These results were obtained both in intact animals (n = 24), which showed ultradian and circadian rhythmicity in behaviour, and in animals (n = 21) with electrolytic lesions aimed at the suprachiasmatic nuclei (SCN), which lacked the circadian modulation of behaviour. Simultaneous deprivation of water and food in 8 voles without circadian rhythmicity during 40 experiments also did not lead to any change in the ultradian periodicity of feeding attempts.Rest deprivation was studied in 5 SCN lesioned voles, by forcing running wheel activity to continue following spontaneous running. Thus, the experimental activity bout was artificially lengthened to 2–9 h in 67 experiments. The onset of the subsequent rest episodes occurred independent of the duration of the preceding . The duration of was dependent on the preceding, experimental in a periodic fashion. The interval experimental (=lengthened +following ) was equal to one, two or three times the control (obtained on nonexperimental days). This result fits the prediction of a clock model and is in conflict with a monotonicincrease of with , as expected in a homeostatic, restorative process.It is concluded that the ultradian timing of activity in the common vole can be explained neither by homeostatic hunger or thirst mechanisms nor by homeostatic rest/activity regulation. The results strongly suggest an independent clock system generating ultradian feeding rhythms in the common vole.Abbreviations
DD
continuous darkness
-
LD
light-dark regime
-
LL
continuous light
-
RCA
retrochiasmatic area
-
ARC
arcuate nucleus
-
SCN
suprachiasmatic nuclei
-
ultradian period
-
ultradian activity time
-
ultradian rest time 相似文献
2.
Summary 2-Amino-2-deoxyuridine reacts efficiently with nucleoside 5-phosphorimidazolides in aqueous solution. The dinucleoside monophosphate analogues were obtained in yields exceeding 80% under conditions in which little reaction occurs with the natural nucleosides.In a similar way, the 5-phosphorimidazolide of 2-amino-2-deoxyuridine undergoes self-condensation in aqueous solution to give a complex mixture of oligomers.The phosphoramidate bond in the dinucleoside monophosphate analogues is stable for several days at room temperature and pH 7. The mechanisms of their hydrolysis under acidic and alkaline conditions are described.Abbreviations A
adenosine
- C
cytidine
- G
guanosine
- U
uridine
- T
thymidine
- UN
3
2-azido-2-deoxyuridine
- UNH
2
2-amino-2-deoxyuridine
- ImpA
adenosine 5-phosphorimidazolide
- ImpU
uridine 5-phosphorimidazolide
- ImpUN
3
2-azido-2-deoxyuridine 5-phosphorimidazolide
- ImpUNH
2
2-amino-2-deoxyuridine 5-phosphorimidazolide
- pA
adenosine 5-phosphate
- pU
uridine 5-phosphate
- pUN
3
2-azido-2-deoxyuridine 5-phosphate
- pUNH
2
2-amino-2-deoxyuridine 5-phosphate
- UpA
uridylyl-[35]-adenosine
- UpU
uridylyl-[35]-uridine
- UNpA
adenylyl-[52]-2-amino-2-deoxy-uridine
- UNpU
uridylyl-[52]-2-amino-2-deoxyuridine (pUN)n n=2,3,4 [25]-linked oligomers of pUNH
2 poly(A) polyadenylic acid
- Im
imidazole
- MeIm
l-methylimidazole 相似文献
3.
Summary Short oligocytidylates can act as templates for the self-condensation of guanosine 5-phosphorimidazolide. In the absence of a catalytic metal ion or in the presence of Pb2+ a noticeable template effect is already observed with the dimer and the yield of long oligomers reaches a plateau with a hexamer template. Short templates give oligomers longers than the template length. The products are predominantly 2-5 linked for the Pb2+-catalyzed reaction while mixed linkages are observed in the uncatalyzed reaction.In the presence of Zn2+, a template effect is first observed with the pentamer and is maximal by the heptamer. The products are predominantly 3-5 linked. Oligomers shorter than or as long as the template are obtained in substantial yield, and longer products in much lower yields.Abbreviations G
Guanosine
- Gp
guanosine 2(3)-phosphate
- pG
guanosine 5-phosphate
- Gp!
guanosine cyclic 2,3-phosphate
- ImpG
guanosine 5-phosphorimidazolide
- ImpG*
[8-14C]-guanosine 5-phosphorimidazolide
- pGp
5-phosphoguanosine 2(3)-phosphate
- G2pG
guanylyl-[2-5]-guanosine
- G3pG
guanylyl-[3-5]-guanosine
- ImpGpG
5-phosphorimidazolide of GpG
- (pG)n (n = 2,3)
oligomers of pG
- GppG
P1, P2-diguanosine 5-diphosphate
- GppGpG
5-[guanosine 5-pyrophosphate] of GpG
- NH2pG
guanosine 5-phosphoramidate
- (pG)4+
tetramer and higher oligoguanylates with 5 terminal phosphate
- oligo(G)
oligoguanylate
- Cp
cytidine 2(3)-phosphate
- Cp!
cytidine cyclic 2,3-phosphate
- (Cp)n–1 Cp! (n= 2,3,4)
oligocytidylates terminated by 5-OH groups and 2,3-cyclic phosphates
- oligo(C)
oligocytidylate
- poly(C)
polycytidylic acid
- poly(U)
polyuridylic acid
- poly(C,G)
random copolymer of C and G
- BAP
bacterial alkaline phosphatase (E. coli)
- EDTA
ethylenediaminetetraacetic acid
- Rf
chromatographic mobility 相似文献
4.
The cleavage of adenosine-5-monophosphate (5-AMP) and guanosine-5-monophosphate (5-GMP) by Ce4+ and lanthanide complex of 2-carboxyethylgermanium sesquioxide (Ge-132) in acidic and near neutral conditions was investigated by NMR , HPLC and measuring the liberated inorganic phosphate at 37°C and 50°C. The results showed that 5-GMP and 5-AMP was converted to guanine (G), 5-monophosphate (depurination of 5-GMP), ribose (depurination and dephosphorylation of 5-GMP), phosphate and adenine (A), 5-monophosphate (depurination of 5-AMP), ribose (depurination and dephosphorylation of 5-AMP), phosphate respectively by Ce4+. In presence of lanthanide complexes, 5-GMP and 5-AMP were converted to guanosine (Guo) and phosphate and adenosine (Ado) and phosphate respectively. The mechanism of cleaving 5-GMP and 5-AMP is hydrolytic scission 相似文献
5.
Summary We have studied the reactions between adenosine 5-phosphorimidazolide and 9-(2-amino-2-deoxyxylofuranosyl) adenine (I) or 3-methylamino-3-deoxyadenosine (II), both with and without a poly (U) template. We find that both amino compounds react much more rapidly than does adenosine, in the absence of a template. The rate of reaction is greatly enhanced by a poly (U) template in the case of I, but the enhancement is slight in the case of II.Abbreviations A
adenosine
- xylo ANH2
9-(2-amino-2-deoxy--D-xylofuranosyl) adenine
- ANHMe
3-methylamino-3-deoxyadenosine
- ImpA
adenosine 5-phosphorimidazolide
- A3 pA
adenylyl-[35]-adenosine
- A2 pA
adenylyl-[25]-adenosine
- UNPA
adenylyl-[52]-2-amino-2-deoxyuridine
- xylo ANPA
9-[adenylyl-(52)-2-amino-2-deoxy--D-xylofuranosyl]adenine
- A(NMe)pA
adenylyl-[53]-3-methylamino-3-deoxyadenosine
- pA
adenosine 5phosphate
- AppA
P1, P2-diadenosine 5pyrophosphate
- (pA)n
n = 2, 3 [2-5]-linked oligomers of pA
- A2 pA2 pA
[2-5]-linked trinucleoside diphosphate of A
- poly (U)
polyuridylic acid 相似文献
6.
Purified ribulose-bisphosphate carboxylase (EC 4.1.1.39) was strongly and equally inhibited either by ADP or GDP and to a lesser extent by IDP. AMP or ATP exerted little effect on activity. Inhibition by the nucleotide diphosphates was competitive with respect to RuBP and non-competitive with respect to CO2 and Mg2+, respectively. Treatment of the enzyme with urea or guanidine-HCl resulted in rapid loss of activity that was not restored by dialysis even in the presence of Mg2+ and cysteine. Sodium dodecyl sulfate electrophoresis of 8.0 M urea treated enzyme revealed the presence of a fast-moving (small) sub-unit with molecular weight 14150 and a slower moving (large) sub-unit with molecular weight 68000. Examination of native enzyme by sodium dodecyl sulfate electrophoresis gave sub-units of 13700 and 55500 respectively. The amino acid content standardized to phenylalanine was essentially similar to that from other sources. Arrhenius plots showed a break at 29°C with an E
a of 12.34 kcal per mole for the steeper part of the curve and a H of 11.43 kcal per mole while for the less steep region, the E
a was 1.04 kcal per mole and the H 1.92 kcal per mole.Abbreviations ADP
adenosine-5-diphosphate
- AMP
adenosine-5-monophosphate
- ATP
adenosine-5-triphosphate
- CDP
cytidine-5-diphosphate
- CMP
cytidine-5-monophosphate
- CTP
cytidine-5-triphosphate
- FDP
fructose-1,6-diphosphate
- F6P
fructose-6-phosphate
- GDP
guanosine-5-diphosphate
- GMP
guanosine-5-monophosphate
- G6P
glucose-6-phosphate
- GTP
guanosine-5-triphosphate
- IDP
inosine-5-diphosphate
- IMP
inosine-5-monophosphate
- PEP
phosphoenolpyruvate
- 6PG
6-phosphogluconate
- R1P
ribose-1-phosphate
- R5P
ribose-5-phosphate
- RuBP
ribulose-1,5-bisphosphate
- SDS
sodium dodecyl sulfate
- TDP
thymidine-5-diphosphate
- TMP
thymidine-5-monophosphate
- TTP
thymidine-5-triphosphate
- UDP
uridine-5-diphosphate
- UMP
uridine-5-monophosphate
- UTP
uridine-5-triphosphate 相似文献
7.
Ahlert Schmidt 《Archives of microbiology》1977,112(3):263-270
Crude extracts of Rhodospirillum rubrum catalyzed the formation of acid-volatile radioactivity from (35S) sulfate, (35S) adenosine-5-phosphosulfate, and (35S) 3-phosphoadenosine-5-phosphosulfate. An enzyme fraction similar to APS-sulfotransferases from plant sources was purified 228-fold from Rhodospirillum rubrum. It is suggested here that this enzyme is specific for adenosine-5-phosphosulfate, because the purified enzyme fraction metabolized adenosine-5-phosphosulfate, however, only at a rate of 1/10 of that with adenosine-5-phosphosulfate. Further, the reaction with 3-phosphoadenosine-5-phosphosulfate was inhibited with 3-phosphoadenosine-5-phosphate whereas this nucleotide had no effect on the reaction with adenosine-5-phosphosulfate. For this activity with adenosine-5-phosphosulfate the name APS-sulfotransferase is suggested. This APS-sulfotransferase needs thiols for activity; good rates were obtained with either dithioerythritol or reduced glutathione; other thiols like cysteine, 2-3-dimercaptopropanol or mercaptoethanol are less effective. The electron donor methylviologen did not catalyze this reaction. The pH-optimum was about 9.0; the apparent K
m for adenosine-5-phosphosulfate was determined to be 0.05 mM with this so far purified enzyme fraction. Enzyme activity was increased with K2SO4 and Na2SO4 and was inhibited by 5-AMP. These properties are similar to assimilatory APS-sulfotransferases from spinach and Chlorella.Abbreviations APS
adenosine-5-phosphosulfate
- PAPS
3-phosphoadenosine-5-phosphosulfate
- 5-AMP
adenosine-5-monophosphate
- 3-AMP
adenosine-3-monophosphate
- 3-5-ADP
3-phosphoadenosine-5-phosphate (PAP)
- DTE
dithiorythritol
- GSH
reduced glutathione
- BAL
2-3-dimercaptopropanol 相似文献
8.
Summary In the human proteolipid protein gene, the base sequence of the intronic region 5 to exon 6 was found to be 5-ctctttcattttcctgcag-3 and not 5-ctctttt-cattttcctgcag-3 as previously reported. 相似文献
9.
Summary Nucleoside-5-triphosphates such as 5-ATP and 5-GTP can be produced efficiently and continuously from 3-mononucleotides such as 3-AMP and 3-GMP by a series of processes consisting of two reaction phases using dried cells of Candida sp. N-25-2 (a hydrocarbon assimilating yeast). Moreover, incidentally to the 5-triphosphates, free uracil is yielded almost stoichiometrically from 3-CMP and 3-UMP which, as is well known, are main concomitant products depolymerized from RNA. Uracil is then also available for many usage. 相似文献
10.
P. Szabo C. Panneerselvam M. Clinton M. Frangou-Lazaridis D. Weksler E. Whittington M. J. Macera K.-H. Grzeschik A. Selvakumar B. L. Horecker 《Human genetics》1993,90(6):629-634
A genomic clone encoding prothymosin (gene symbol: PTMA), a nuclear-targeted protein associated with cell proliferation, was isolated and the 5-regulatory region subcloned and sequenced. Because of previously reported discrepancies between several cDNA clones and a genomic clone for prothymosin , we determined the sequence of the first exon and of a 1.7-kb region 5 to the first exon. The sequence of the genomic clone reported here corresponds to the published cDNA sequences, suggesting that the previously noted discrepancies may be due to genetic polymorphism in this region. In addition, our sequence data extend the known 5-upstream sequence by an additional 1.5 kb allowing the identification of numerous, potential cis-acting regulatory sites. This 5-flanking cloned probe permitted us to localize the protothymosin gene to chromosome 2 in humans. 相似文献
11.
Summary The self-condensation of 2(3)-O-glycyl esters of adenosine, adenosine-5-(O-methylphosphate) and P1, P2-diadenosine-5-pyrophosphate in 6.2 mM solutions at pH 8.0 and -5°C in the presence of 12.5 mM poly(U) yields approximately 3 times as much diketopiperazine as reactions without poly(U). As the concentration of 2(3)-O-(glycyl)-P1, P2-diadenosine-5-pyrophosphate is decreased from 6.2 mM to 1.5 mM the yield of diketopiperazine in the presence of poly(U) decreases slightly from 6.6% to 5.2%, whereas, in the absence of poly(U) the yield of diketopiperazine decreases substantially from 2.4% to 0.75%. The enhanced yield of diketopiperazine that is attributed to the template action of poly(U) is temperature dependent and is observed only at temperatures below 10°C (5°C to -5°C) for 6.2 mM 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate) and below 23°C (15°C to -5°C) for 6.2 mM 2(3)-O-(glycyl)-P1, P2-diadenosine-5-pyrophosphate. The absence of a template effect at high temperatures is attributed to the melting of the organized helices. The hydrolysis half-lives at pH 8.0 and -5°C of 2(3)-O-(glycyl)-adenosine, 2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate), 2(3)-O-(glycyl)-P1, P2-diadenosine-5-pyrophosphate, and 5-O-(glycyl)-adenosine in the presence of poly(U) are substantially larger than their half-lives in the absence of poly(U). The condensation of 2(3)-O-(glycyl)-adenosine yields 5% of 5-O-(glycyl)-adenosine in the presence of poly(U) compared to 0.7% in the absence of poly(U).Abbreviations DKP
diketopiperazine
- (gly)2
glycylglycine
- (gly)3
glycylglycylglycine
- AppA-gly
2(3)-O-(glycyl)-P1, P2-diadenosine-5-pyrophosphate
- MepA-gly
2(3)-O-(glycyl)-adenosine-5-(O-methylphosphate)
- Ado-2(3)-gly
2(3)-O-(glycyl)-adenosine
- Ado-5-gly
5-O-(glycyl)-adenosine
- Boc-gly
N-tert-butyloxycarbonylglycine
- AppA
P1, P2-diadenosine-5-pyrophosphate
- MepA
adenosine-5-(O-methylphosphate)
- AppA-Boc-gly
2(3)-O-(Boc-glycyl)-P1, P2-diadenosine-5-pyrophosphate
- Ado-5-Boc-gly
5-O-(Boc-glycyl)-adenosine
- Ado-2(3)-Boc-gly
2(3)-O-(Boc-glycyl)-adenosine 相似文献
12.
Adenosine-5-triphosphate was synthesized by the phosphorylation of adenosine-5-diphosphate in aqueous solution containing cyanate as a condensing reagent and insoluble calcium phosphate produced from phosphate and calcium chloride. In a similar manner, adenosine-5-diphosphate was synthesized from adenosine-5-monophosphate. When the experiment was carried out in the conditions of 4 °C and pH 5.75, the formation of adenosine-5-diphosphate and adenosine-5-triphosphate from adenosine-5-monophosphate was observed in the yields of 19 and 7%, respectively. The other nucleoside-5-triphosphates were also produced from their respective diphosphates. 相似文献
13.
Summary Evaporation of a solution of thymidine plus either theexo or theendo diastereomer of uridine cyclic 2,3-O, O-phosphorothioate (U > p(S) in 1,2-diaminoethane hydrochloride buffer gave the 2,5 and 3,5 isomers of (P-thio) uridylylthymidine (Up(S)dT) in a ratio of 1:2 with a combined yield of about 20%. These isomers were re-converted to U > p(S) and dT by a reaction that is known to proceed by an in-line mechanism. Both the 2,5 and 3,5 isomers gave as product the same diastereomer of U > p(S) that had been used originally in their formation. These dry-state prebiotic reactions (Verlander, Lohrmann, and Orgel 1973) are thus shown to be stereospecific, and both the 2,5 and 3,5 internucleotide bonds are formed by an in-line mechanism.Abbreviations DAE
1,2-diaminoethane
- HPLC
high pressure liquid chromatography
- RNase
bovine pancreatic ribonuclease A, EC 3.1.4.22
- TEAB
triethylammonium bicarbonate
- tris
tris(hydroxymethyl)aminomethane
- UMP(S)
uridine monophosphorothioate
- U > p
uridine cyclic 2,3-phosphate
- U > p(S)
uridine cyclic 2,3-O, O-phosphorothioate
- Up(S)dT
(P-thio)uridylylthymidine
- U2p(Rp-S)5dT
(P-thio)uridylylthymidine with theR configuration at phosphorous, and a 2,5 internucleotide linkage 相似文献
14.
The six binary montmorillonite clay-catalyzed reactions of the5-phosphorimidazolides of adenosine, cytidine, guanosine anduridine were performed and the eight dimers from each reactionwere separated and analyzed by HPLC. A 16–51-fold higher yieldof the 5-purine-pyrimidine dimers over that of the5-pyrimidine-purines was observed. The total yield of the5-purine-pyrimidine dimers was in the 50–70% range while thatof the 5-pyrimidine-purine dimers was 1.3–7.0%. Less sequenceselectivity was observed in the homodimers formed.Regioselectivity for the formation of 3, 5-phosphodiesterbonds over that found in the absence of clay was observed. The5-purine-pyrimidine, 5-pyrimidine-pyrimidine and5-purine-purine dimers had 3, 5-links in about half of theirphosphodiester bonds. The percent phosphodiester links in the5-pyrimidine-pyrimidine dimers was 18%, a value close to thatobserved in the absence of the montmorillonite catalyst. Themontmorillonite-catalyzed reaction of all four activatednucleotides was performed and the 24 products were separated andanalyzed. The trends observed in the binary reactions wereconfirmed and the results also showed that the relativereactivity of the activated monomers was A>G>C>U in theratio 8.2: 4.8: 1.3: 1 respectively. No 5-pyrimidine-purineswith a 5-U and pG3pU, pC3pAand pC3pG weredetected. These studies suggest that a limited population ofRNAs would have formed in catalyzed prebiotic reactions. 相似文献
15.
Summary Soluble lead salts and a number of lead-containing minerals catalyze the formation of oligonucleotides from nucleoside 5-phosphorimidazolides. The effectiveness of lead compounds correlates strongly with their solubility. Under optimal conditions we were able to obtain 18% of pentamer and higher oligomers from ImpA. Reactions involving ImpU gave smaller yields.Abbreviations A
adenosine
-
U
uridine
-
Im
imidazole
-
MeIm
1-methyl-imidazole
-
EDTA
ethylenediaminetetraacetic acid
-
pA
adenosine 5-phosphate
-
pU
uridine 5-phosphate
-
Ap
adenosine cyclic 2:3-phosphate
-
ATP
adenosine 5-triphosphate
-
AppA
P1,P2-diadenosine 5-diphosphate
-
pNp (N = A,U)
nucleotide 2(3), 5-diphosphate
-
ImpA
adenosine 5-phosphoreimidazolide
-
ImpU
uridine 5-phosphorimidazolide
-
A
2pA
adenylyl-[25]-adenosine
-
A
3pA
adenylyl-[35]-adenosine
-
pA
2pA
5-phospho-adenylyl-[25]-adenosine
-
pA
3pA
5-phospho-adenylyl-[35]-adenosine
-
pUpU
5-phospho-uridylyl-uridine
-
pApU
5-phospho-adenylyl-uridine
-
pUpA
5-phospho-uridylyladenine
-
(pA)n (n, 2,3,4,)
oligoadenylates with 5 terminal phosphate
-
ImpApA
5-phosphorimidazolide of adenylyl adenosine
-
(pA)
5+
pentamer and higher oligoadenylates with 5 terminal phosphate
-
(Ap)nA (n = 2,3,4)
oligoadenylates without terminal phosphates
In the following we do not specify the nature of the internucleotide linkageIn the following we do not specify the nature of the internucleotide linkage 相似文献
16.
17.
18.
Summary We have studied a number of condensation reactions involving ImpU, ImpT, ImpC, ImpA, ImpG, ImpUpG and ImpCpA as activated nucleotide donors and a variety of homo- and hetero-polynucleotides as templates. We did not obtain any evidence of a template effect with ImpU and ImpT, but observed some condensation of ImpC with GpG on appropriate templates. ImpA and ImpG take part in a number of more or less efficient template-directed reactions, as do ImpUpG and ImpCpA.Our results suggest that, on the primitive Earth, pyrimidine nucleotides could most easily have been incorporated into polymers as constituents of short oligomers, which contained one or more purine nucleotide. The linkage of the product depends strongly on the nature of the substrates; the percentage of the natural 3-5-linkage was, in some cases, less than 10% and, in others, as high as 70%. Wobble-pairing was often very effective in promoting condensations, suggesting that transition mutations would have been very frequent in prebiotic polynucleotide replication.Abbreviations and Conventions U
uridine
- T
thymidine
- C
cytidine
- A
adenosine
- G
guanosine
- pN
nucleoside-5-phosphate
- Np
a mixture of 2- and 3-phosphates of a nucleoside
- pNp
a mixture of the 2-5-diphosphate and 3-5-diphosphate of a nucleoside
- N1
2 pN2
a 2-5-linked dinucleoside monophosphate
- N1
3 pN2
a 3-5-linked dinucleoside monophosphate
- N5 ppN
a pyrophosphate derived from a nucleoside-5-phosphate. ImpN and ImpN1pN2 are 5-phosphorimidazolides of nucleosides and 3-5-linked dinucleoside monophosphates, respectively
- poly(N)
a homopolynucleotide
- poly (U1 C2 A4 G3)
a random copolymer derived from a substrate mixture containing U, C, A, G in ratio 1:2:4:3
- ODU
optical density units measured at 260 nm 相似文献
19.
Adenine deoxynucleotides bind more strongly to Na+-montmorillonite than do the corresponding ribonucleotides. Thymidine nucleotides binds less strongly to Na+-montmorillonite than do the corresponding adenine deoxynucleotides. Oligomers of 2-dpA up to the tetramer were detected in the reaction 2-d-5-AMP with EDAC (a water-soluble carbodiimide) in the presence of Na+-montmorillonite. Reaction of 3-d-5-AMP with EDAC on Na+-montmorillonite yields 3-d-2,5-pApA while the reaction of 2-d-3-AMP yields almost exclusively 3,5-cdAMP. The reaction of 5-TMP under the same reaction conditions give 3,5-cpTpT and 3,5-pTpT while 3-TMP gives mainly 3,5-cpT. The yield of dinucleotide products (dpNpN) containing the phosphodiester bond is 1% or less when Na+-montmorillonite is omitted from the reaction mixture. 相似文献
20.
Purified myelin from fresh calf brain white matter was subfractionated in a discontinuous sucrose gradient; significant recovery of protein and 2,3-cyclic nucleotide 3-phosphohydrolase (CNP) and 5-nucleotidase (5N) activities occurred in all three obtained subfractions, the highest recovery being in the light subfraction; highest 5N and CNP specific activities were in medium myelin. Purified myelin was also subfractionated in a continuous sucrose gradient, with a similar localization of protein; CNP activity and 5N activity maxima suggest that myelin may be a predominant locus of 5N in bovine brain white matter. Freezing of brain white matter caused an increase in protein and in CNP and 5N total activity recoveries in denser myelin subfractions. Cytochemistry showed the reaction product of 5N in the whole myelin fraction to be associated with the innermost, outermost and medial compact myelin layers. Effects of non-ionic detergent (Lubrol WX) on 5N activity were studied, and the results also suggest the intrinsic nature of 5N as an ectoenzyme in myelin membranes. Lubrol WX was viewed as an advisable detergent for the stimulation of myelin 5N activity, but not for the solubilization of this enzyme. 相似文献