Purification and kinetic analysis of a baculovirus ecdysteroid UDP-glucosyltransferase

EVANS, O. P.; O'REILLY, D. R.

doi:10.1042/bj3320807u

Cited by 12 publications

(20 citation statements)

References 17 publications

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“…12fNo. 3 UDP-glycosyltransferases catalyzing the addition of a glycosyl moiety from an UDP-sugar to an aglycone, such as T 4 phage (3-glucosyltransferase (J 3), baculovirus ecdysteroid UDP-glucosyltransferase (14), and flavonol 3-0-galactosyltransferase and limonoid UDPglucosyltransferase in plants (15,16), are known as (3-glycosyltransferases . Further characterization of BGluT will be of interesting in the point of its catalytic mechanism and evolutionary relationship between other (3-g1ycosyJ types of pteridine glycosyltransferases.…”

Section: Resultsmentioning

confidence: 99%

Chemical structure of 1-O-(L-erythro-biopterin-2'-yl)-a-glucose isolated from a cyanobacterium Synechococcus sp. PCC 7942

et al. 2001

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A pteridine glycoside in Synechococcus sp. PCC 7942, the structure of which had been tentatively identified as biopterin-glucoside, was isolated and characterized for its exact chemical structure by 2D-NMR spectroscopy. The determined structure is 1-0-(L-erythro-biopterin-2'-yl)-a-glucose. It is the first report on the occurrence of a biopterin-glucoside having a a-configured sugar directly attached at the pteridine ring. This result also supports that the previously purified UDP-glucose: BH4 glucosyltransferase from Synechococcus sp. PCC 7942, which catalyzes the synthesis of BH4-glucoside from UDP-glucose and BH4, is a a-glucosyltransferase.

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Section: Resultsmentioning

confidence: 99%

Chemical structure of 1-O-(L-erythro-biopterin-2'-yl)-a-glucose isolated from a cyanobacterium Synechococcus sp. PCC 7942

et al. 2001

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“…2F). This observed K cat of the purified E22O for ecdysone was much higher than those of the ecdysteroid UDP-glucosyltransferase of the Autographa californica nucleopolyhedrovirus (K cat ϭ 0.069/s) and ecdysone oxidase of the cotton leafworm Spodoptera littoralis (K cat ϭ 0.11-0.12/s, calculated assuming that the molecular mass of the enzyme is 190 kDa), whereas the K m values of all three enzymes for ecdysone were similar (19,33). These results suggest that E22O inactivates ecdysone much more efficiently than the two other well characterized ecdysteroid inactivation enzymes.…”

Section: Purification Kinetic Analysis and Cdna Cloning Of E22o-mentioning

confidence: 99%

Fungal Ecdysteroid-22-oxidase, a New Tool for Manipulating Ecdysteroid Signaling and Insect Development

Kamimura

Saitô

Niwa

et al. 2012

Journal of Biological Chemistry

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Background: Artificial reduction of internal ecdysteroid titer is very difficult to achieve in insects. Results: Injection of Nomuraea rileyi ecdysteroid-22-oxidase (E22O) or forced expression of the E22O gene reduced ecdysteroid titer and manipulated embryogenesis, molting, metamorphosis, and diapause in a number of insects. Conclusion: E22O is the first versatile ecdysteroid titer-decreasing tool. Significance: E220 will be used to answer various ecdysteroid-associated developmental and physiological questions.

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“…EGT activity was assayed as described in Evans & O'Reilly (1998). Briefly, EGT was incubated at 37 mC for 30 min in the presence of 20 µM ecdysone, 0n05 µCi [$H]UDP-glucose and 10 mM MgCl # (unless noted otherwise).…”

Section: Methodsmentioning

confidence: 99%

“…Enzyme action on ecdysone and UDP-glucose results in the formation of ecdysone-22-O-β--glucoside . It can accept sugars from either UDP-glucose or UDPgalactose with almost equal efficiency in vitro (Evans & O'Reilly, 1998). However, only ecdysteroid galactosides are formed during AcMNPV infection in vivo, presumably because UDP-galactose is the predominant UDP-sugar available to the enzyme in the insect host Evans & O'Reilly, 1998).…”

Section: Introductionmentioning

confidence: 99%

“…AcMNPV EGT can conjugate a range of ecdysteroids provided they possess a hydroxyl group at position C-22 . However, it displays very different specificities for different ecdysteroids showing, in particular, a marked preference for ecdysone and 3-dehydroecydsone over 20-hydroxyecdysone, the most active form of the hormone in vivo (Evans & O'Reilly, 1998).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Expression and structural characterization of a baculovirus ecdysteroid UDP-glucosyltransferase.

Evans

O’Reilly

1999

Journal of General Virology

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The baculovirus enzyme ecdysteroid UDP-glucosyltransferase (EGT) disrupts the hormonal balance of the insect host by catalysing the conjugation of ecdysteroids, the moulting hormones, with the sugar moiety from UDP-glucose or UDP-galactose. In this study, EGT has been overproduced using a recombinant Autographa californica nucleopolyhedrovirus and an antiserum has been raised against the purified protein. This antiserum was used to visualize the kinetics of expression of EGT by wild-type AcMNPV L-1 and by the overproducing recombinant virus. The inclusion of tunicamycin during these time-course experiments suggested that EGT is glycosylated. This was confirmed by Endo F treatment, which showed that glycosylation increased the apparent subunit molecular mass by approximately 11 kDa. These sugars do not appear to be required for enzyme activity. EGT activity invariantly elutes from gel-filtration columns as a single peak corresponding to a 260 kDa (O50 kDa) protein. This suggests that the enzyme is an oligomer of three to five subunits, since the subunit molecular mass is 56 kDa.

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Purification and kinetic analysis of a baculovirus ecdysteroid UDP-glucosyltransferase

Cited by 12 publications

References 17 publications

Chemical structure of 1-O-(L-erythro-biopterin-2'-yl)-a-glucose isolated from a cyanobacterium Synechococcus sp. PCC 7942

Chemical structure of 1-O-(L-erythro-biopterin-2'-yl)-a-glucose isolated from a cyanobacterium Synechococcus sp. PCC 7942

Fungal Ecdysteroid-22-oxidase, a New Tool for Manipulating Ecdysteroid Signaling and Insect Development

Expression and structural characterization of a baculovirus ecdysteroid UDP-glucosyltransferase.

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