Studies of the phosphodiesterase from rattlesnake venom

Garilhe, M. Privat de; Laskowski, M.

doi:10.1016/0006-3002(55)90100-1

Cited by 85 publications

(10 citation statements)

References 15 publications

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“…Razzell and Khorana (1 4,15), during extensive studies of the hydrolysis of low molecular weight substrates found that although venom phosphodiesterase action was primarily exonucIeolytic, there was, in agreement with similar findings by Laskowski et a]. (16,17), a detectable degree of endonucleolytic hydrolysis with certain substrates. It was of interest to determine the extent of endonucleolysis of highly polymerized RNA under standardized conditions since the assumption of an exclusively exonudeolytic attack by phosphodiesterase on RNA Is frequently made and has important implications in regard to the interpretation of experimental findings.…”

supporting

confidence: 80%

The chain termini of polynucleotides formed by limited enzymic fragmentation of wheat embryo ribosomal RNA. Part 1. Studies of snake venom phosphodiesterase

McLennan¹,

Lane²

1968

Can. J. Biochem.

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The chain termini of polynucleotides formed by limited enzymic fragmentation of wheat embryo ribosomal RNA. Part 1. Studies of snake venom phosphodiesterase. Can. J. Biochem. 46, 81 (1968).Snake venom phosphodiesterase induces about fifteen exonucleolytic cleavages for each endonucleolytic cleavage during the first hour of hydrolysis sf wheat embryo ribosomal RNA, under the conditions of hydrolysis used in this present investigation. The polynucleotide chains in the sibosornaI RNA preparation have am average degree of polymerkation in the neighborhood of 1308 nucleotide residues, and there is a mean of between 5 md 18 endonucleolytic breaks per chain during this airst hour of phosphodiesterase-induced hydrolysis. The cleavages occur widely throughout most of the polynucleotide chains in the ribosomal RNA preparation, as judged by the sharp decrease in mean sedimentation rate which accompanies a limited degree (about 10%) of exonucleolpis of the RNA. Studies of phosphodiestaase-indumd endonucleolysis of wheat embryo soluble RNA are reported, but because of the much lower initial degree of polymerization (about 88 nucleotide residues per polynucleotide chain), the results of endonucleolysis are less pronounced in terms of the proportional increment in chain termini and the proportional decrease of mean sedimentation sate. The endonucleolysis of RNA is discussed in tams of the minor nucleotide components in both ribosomal and soluble RNA, and particular reference is made to pseudouridylate which has been found in relatively high proportion among the chain termini after limited hydrolysis with venom phosphodiesterase.Purified venom phosphodiesterase preparations, devoid of ribonuclease or 5'-naacleotidase contamination, were found to convert nucleoside 2'(3'),5'-diphosphsates to 5'-nucleotides under conditions which had virtually no effect on nucleoside 3'-phosphates or nucleoside %'-phosphates. The possibility that this reaction may be catalyzed by the venom phosphodiesterase itself is discussed.In 1951, Marko and Butler (1) introduced the sodium dodecyl suKate procedure for the isolation of calf thymus DNA', and in the same year, Hurst and Butler (2), succeeded in obtaining snake venom phosphodiesterase preparations, which in concert with pancreas DNase effected an almost quantitative conversion of the highly polymerized DNA to its constituent 5'-deoxyr-ibonucleotides (4). By 1955, Helleiner (4) had established that the hydrolysis catalyzed by venom phosphodiesterase was exonucieoiyiic. Although this exonucleolytic action of venom phosphodiesterase has been exploited to advantage,there have been few studiesdesiped to assess the extent to which venom phosphodiesterase 'Abbreviations : DNA, deoxyribonucleate(s) ; rRNA, ribosomal ribonuclea%e(s) ; sRNA, solublle ribsnucleate(s) ; NxpNp, alkali-stable dinucleotides containing 2'-Qmthylribose in which N is any of the four major riboaaucleosides in RNA and Nx is the 2'-0-methylated derivative of m y sf the four principal nuclmsides; pA, adenosine 5'-phosphate; A2p, adeno...

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confidence: 80%

The chain termini of polynucleotides formed by limited enzymic fragmentation of wheat embryo ribosomal RNA. Part 1. Studies of snake venom phosphodiesterase

McLennan¹,

Lane²

1968

Can. J. Biochem.

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“…The use of deoxyribonuclease 11 (spleen acid deoxyribonuclease), which has a pHoptimum near pH 5.0 [53,54], gave less reproducible results, because it produces more of the higher oligonucleotides than deoxyribonuclease I [55].…”

Section: Purification Of D-galactose Dehydrogenasementioning

confidence: 99%

d‐Galactose Dehydrogenase from Pseudomonas fluorescens

Blachnitzky

Wengenmayer

Kurz

1974

European Journal of Biochemistry

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1. A procedure is described for the isolation of D-galactose dehydrogenase from Pseudomonas fluorescens, grown on D-galactose as carbon source. A 750-fold purification was achieved with an overall yield of about 40%. The final preparation had a specific activity of about 850 pmo1 NADH formed per min per mg protein.2. The enzyme has been shown to be homogeneous by ultracentrifugation, by disc gel electrophoresis, by gel isoelectric focusing and by dodecylsulfate gel electrophoresis.3. The molecular weight of the native enzyme has been determined to be 64000 by sedimentation equilibrium studies and by gel permeation chromatography.4. The enzyme dissociates in the presence of 0.1 % sodium dodecylsulfate to polypeptide chains, whose molecular weight was determined as 32000 by electrophoresis, indicating that the native enzyme is composed of two subunits.5. The amino acid composition of D-galactose dehydrogenase has been determined. 6. The PI of the enzyme has been shown to be 4.28. The pH optimum is between 9.1 and 9.5. At pH 9.1 and 30 "C the limiting Michaelis constant for NAD+ is Ka = 0.24 mM, for NADP+ Ka = 2.3 mM and for D-galactose with NAD+ and with NADP+ as cosubstrate Kb = 0.7 mM. The dissociat on constant for NAD+ is Kia = 0.54 mM and for NADPI-Kia = 6.2 mM. 8. The correspondence between amino acid composition, subunit size and structure and between specificity of D-galactose dehydrogenase from Ps. fluorescens and from Ps. saccharophila is discussed.

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“…T h e standard deviations of the values for replicate determinations on the same dinucleotide sample were d= 1-2% it1 most cases. The molar quantities of the dinuclcotides were estimated by using the sum of the molar extinction coefficients of the constituent nucleosides with no correction having been made for the Iiypochromicity of dinucleotides(21,22). which is arbitrarily assigned a value of 1.00.…”

mentioning

confidence: 99%

The Alkali-Stable Dinucleotide Sequences in 18 s + 28 s Ribonucleates From Wheat Germ

Singh¹,

Lane²

1964

Can. J. Biochem.

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The 2'-0-i~lethyl substitution of ribose residues of ribonucleate chains can give rise to 16 alkali-stable dinucleotides having the general structure NxpNp in which N is any of the four major ribonucleosides and Nx is the 2'-0-methj.1 derivative of any of the four major ribonucleosides. Quantitative data are presented which show that all of these 16 alkali-stable dinucleotides occur in characteristic a~~~o u n t s in alkali hydrolyzates of the 18 S + 28 S ribonucleates from wheat germ. 'The alltali-stable dinucleotitles cumulatively account for 3 mole% of the total nucleotides of the 18 S + 28 S wheat germ ribonucleates. The relative amounts of the four 2'-0-methyl ribosides responsible for the alkali stabilit). of the tlinucleotitlcs have been shown to be distinctly different froin the relative an~ounts of the four corresponding normal ribosides which participate in either the allcali-stable dinucleotide sequences or in the ribonucleate chains as a whole. Quantitati\-e end group data show that the alkali-stable dinucleotide sequences occur internally in high molecular weight ribonucleate chains. Preliminary data are presented 011 the occurrence of alkali-stable trinucleotides in alkali hydrolyzates of the 18 S + 28 S wheat germ ribonucleates.

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Studies of the phosphodiesterase from rattlesnake venom

Cited by 85 publications

References 15 publications

The chain termini of polynucleotides formed by limited enzymic fragmentation of wheat embryo ribosomal RNA. Part 1. Studies of snake venom phosphodiesterase

The chain termini of polynucleotides formed by limited enzymic fragmentation of wheat embryo ribosomal RNA. Part 1. Studies of snake venom phosphodiesterase

d‐Galactose Dehydrogenase from Pseudomonas fluorescens

The Alkali-Stable Dinucleotide Sequences in 18 s + 28 s Ribonucleates From Wheat Germ

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