Purification of NAD glycohydrolase from Agkistrodon acutus venom

Wu, Shuang; Li, Yanli; Xu, Xiaolong; Zhu, Zhen‐Gang

doi:10.1016/s1046-5928(02)00015-3

Cited by 6 publications

(11 citation statements)

References 29 publications

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“…NADase activity was assayed by spectrophotometric measures of decrement of ␤-NAD ϩ , according to Wu et al, 42 with minor modifications. Briefly, the reaction mixture contained 0.05 M potassium phosphate buffer (pH 7.3), 5 mM ␤-NAD, 1 mM MgCl 2 and 200 g of either crude or purified disc preparations in a total volume of 1 mL.…”

Section: Assay For Nad ؉ Glycohydrolase Activitymentioning

confidence: 99%

Localization of the Cyclic ADP-Ribose-Dependent Calcium Signaling Pathway in Bovine Rod Outer Segments

Panfoli¹,

Ravera²,

Fabiano³

et al. 2007

Invest. Ophthalmol. Vis. Sci.

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Section: Assay For Nad ؉ Glycohydrolase Activitymentioning

confidence: 99%

Localization of the Cyclic ADP-Ribose-Dependent Calcium Signaling Pathway in Bovine Rod Outer Segments

Panfoli¹,

Ravera²,

Fabiano³

et al. 2007

Invest. Ophthalmol. Vis. Sci.

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“…AA‐NADase is different from other NADases, because it contains Cu(II) ion and its two chains are linked with disulfide‐bonds 8, 9. DTT, GSH and TCEP can reduce both disulfide bonds and Cu(II) in AA‐NADase as determined by SDS‐PAGE and EPR spectroscopy (Figures 2 and 5).…”

Section: Discussionmentioning

confidence: 99%

“…The purification of AA‐NADase was performed as previously described 9. Protein purity was confirmed using SDS‐PAGE and the concentration of AA‐NADase was calculated from the absorption coefficient ( A

_{1 cm}^{1%}

= 0.66) at 280 nm and the relative molecular weight ( M r = 100 kDa).…”

Section: Methodsmentioning

confidence: 99%

“…AA‐NADase is a unique NADase isolated from the venom of Agkistrodon acutus , because among all identified NADases, only AA‐NADase has disulfide‐bond linkages between two peptide chains and contains Cu(II) ion which is essential for catalyzing the hydrolysis of NAD 8–10. Recently, we have reported that AA‐NADase is a multicatalytic enzyme with both NADase and AT(D)Pase‐like activities, because it not only cleaves the C‐N glycosyl bond of NAD to produce ADPR, but also cleaves the P‐O‐P bond of ATP, ADP and AMP‐PNP to produce AMP 11.…”

Section: Introductionmentioning

confidence: 99%

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Small‐molecule reductants inhibit multicatalytic activity of AA‐NADase from Agkistrodon acutus venom by reducing the disulfide‐bonds and Cu(II) of enzyme

Zhang

et al. 2009

Biopolymers

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AA-NADase from Agkistrodon acutus venom is a unique multicatalytic enzyme with both NADase and AT(D)Pase activities. Among all identified NADases, only AA-NADase contains Cu(II) and has disulfide-bond linkages between two peptide chains. The effects of the reduction of the disulfide-bonds and Cu(II) in AA-NADase by small-molecule reductants on its NADase and ADPase activities have been investigated by polyacrylamide gel electrophoresis, high performance liquid chromatography, electron paramagnetic resonance spectroscopy and isothermal titration calorimetry. The results show that AA-NADase has six disulfide-bonds and fifteen free cysteine residues. L-ascorbate inhibits AA-NADase on both NADase and ADPase activities through the reduction of Cu(II) in AA-NADase to Cu(I), while other reductants, dithiothreitol, glutathione and tris(2-carboxyethyl)phosphine inhibit both NADase and ADPase activities through the reduction of Cu(II) to Cu(I) and the cleavage of disulfide-bonds in AA-NADase. Apo-AA-NADase can recover its NADase and ADPase activities in the presence of 1 mM Zn(II). However, apo-AA-NADase does not recover any NADase or ADPase activity in the presence of 1 mM Zn(II) and 2 mM TCEP. The multicatalytic activity relies on both disulfide-bonds and Cu(II), while Cu(I) can not activate the enzyme activities. AA-NADase is probably only active as a dimer. The inhibition curves for both ADPase and NADase activities by each reductant share a similar trend, suggesting both ADPase and NADase activities probably occur at the same site. In addition, we also find that glutathione and L-ascorbate are endogenous inhibitors to the multicatalytic activity of AA-NADase.

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“…Yost & Anderson (1981) characterized the NADase from Bungarus fasciatus venom, and reported that it was a homodimeric glycoprotein of 120–130,000 Da, having a monomeric molecular weight of 62,000 (denaturing SDS PAGE). The enzyme comprised approximately 0.1% of Bungarus fasciatus venom by mass (Yost & Anderson, 1981; Anderson, Yost & Anderson, 1986) while a value of 0.5% was reported from Deinagkistrodon acutus venom (Wu et al, 2002) using a simpler chromatographic procedure with more sophisticated resins.…”

Section: Introductionmentioning

confidence: 99%

Snake venom NAD glycohydrolases: primary structures, genomic location, and gene structure

Koludarov

Aird

2019

PeerJ

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NAD glycohydrolase (EC 3.2.2.5) (NADase) sequences have been identified in 10 elapid and crotalid venom gland transcriptomes, eight of which are complete. These sequences show very high homology, but elapid and crotalid sequences also display consistent differences. As in Aplysia kurodai ADP-ribosyl cyclase and vertebrate CD38 genes, snake venom NADase genes comprise eight exons; however, in the Protobothrops mucrosquamatus genome, the sixth exon is sometimes not transcribed, yielding a shortened NADase mRNA that encodes all six disulfide bonds, but an active site that lacks the catalytic glutamate residue. The function of this shortened protein, if expressed, is unknown. While many vertebrate CD38s are multifunctional, liberating both ADP-ribose and small quantities of cyclic ADP-ribose (cADPR), snake venom CD38 homologs are dedicated NADases. They possess the invariant TLEDTL sequence (residues 144–149) that bounds the active site and the catalytic residue, Glu228. In addition, they possess a disulfide bond (Cys121–Cys202) that specifically prevents ADP-ribosyl cyclase activity in combination with Ile224, in lieu of phenylalanine, which is requisite for ADPR cyclases. In concert with venom phosphodiesterase and 5′-nucleotidase and their ecto-enzyme homologs in prey tissues, snake venom NADases comprise part of an envenomation strategy to liberate purine nucleosides, and particularly adenosine, in the prey, promoting prey immobilization via hypotension and paralysis.

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Purification of NAD glycohydrolase from Agkistrodon acutus venom

Cited by 6 publications

References 29 publications

Localization of the Cyclic ADP-Ribose-Dependent Calcium Signaling Pathway in Bovine Rod Outer Segments

Localization of the Cyclic ADP-Ribose-Dependent Calcium Signaling Pathway in Bovine Rod Outer Segments

Small‐molecule reductants inhibit multicatalytic activity of AA‐NADase from Agkistrodon acutus venom by reducing the disulfide‐bonds and Cu(II) of enzyme

Snake venom NAD glycohydrolases: primary structures, genomic location, and gene structure

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