Characterization and expression of Arabidopsis UDP-sugar pyrophosphorylase

Litterer, Lynn A.; Schnurr, Judy A.; Plaisance, Kathryn L.; Storey, Kathleen K.; Gronwald, John W.; Somers, David A.

doi:10.1016/j.plaphy.2006.04.004

Cited by 59 publications

(67 citation statements)

References 37 publications

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“…This hypothesis is consistent with the high K m value for UTP. As expected, the affinity and turnover for UDP-L-Ara and UDPGalA are low in agreement with the observation that these nucleotide sugars are poorer substrates (Tables 1 and 2) (19,20,32). Thus, the absence of the C6 hydroxyl group and particularly its substitution leads to a drastic reduction of the enzyme efficiency.…”

Section: Major Usp Follows Simple Michaelis-menten Kinetics-supporting

confidence: 88%

Leishmania UDP-sugar Pyrophosphorylase

Damerow

Lamerz

Haselhorst

et al. 2010

Journal of Biological Chemistry

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The Leishmania parasite glycocalyx is rich in galactosecontaining glycoconjugates that are synthesized by specific glycosyltransferases that use UDP-galactose as a glycosyl donor. UDP-galactose biosynthesis is thought to be predominantly a de novo process involving epimerization of the abundant nucleotide sugar UDP-glucose by the UDP-glucose 4-epimerase, although galactose salvage from the environment has been demonstrated for Leishmania major. Here, we present the characterization of an L. major UDP-sugar pyrophosphorylase able to reversibly activate galactose 1-phosphate into UDP-galactose thus proving the existence of the Isselbacher salvage pathway in this parasite. The ordered bisubstrate mechanism and high affinity of the enzyme for UTP seem to favor the synthesis of nucleotide sugar rather than their pyrophosphorolysis. Although L. major UDP-sugar pyrophosphorylase preferentially activates galactose 1-phosphate and glucose 1-phosphate, the enzyme is able to act on a variety of hexose 1-phosphates as well as pentose 1-phosphates but not hexosamine 1-phosphates and hence presents a broad in vitro specificity. The newly identified enzyme exhibits a low but significant homology with UDP-glucose pyrophosphorylases and conserved in particular is the pyrophosphorylase consensus sequence and residues involved in nucleotide and phosphate binding. Saturation transfer difference NMR spectroscopy experiments confirm the importance of these moieties for substrate binding. The described leishmanial enzyme is closely related to plant UDP-sugar pyrophosphorylases and presents a similar substrate specificity suggesting their common origin.

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Section: Major Usp Follows Simple Michaelis-menten Kinetics-supporting

confidence: 88%

Leishmania UDP-sugar Pyrophosphorylase

Damerow

Lamerz

Haselhorst

et al. 2010

Journal of Biological Chemistry

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“…Thus, two main activities can be proposed for the production of this sugar nucleotide. Either an enzyme with UTP-glucose-1-phosphate uridylyltransferase activity (EC 2.7.7.9) presents a weak galactose-1-phosphate uridylyltransferase activity (EC 2.7.7.10), as described in mammals (55,56), or a broad substrate range UDP-sugar pyrophosphorylase (EC 2.7.7.64) is present in the genome of P. falciparum, as described in plants or Leishmania major (57)(58)(59)(60)(61). UDP-sugar pyrophosphorylase is an enzyme that can nonspecifically utilize UTP and glucose 1-phosphate or galactose 1-phosphate to produce UDP-glucose or UDP-galactose and pyrophosphate.…”

Section: Discussionmentioning

confidence: 99%

Biosynthesis of GDP-fucose and Other Sugar Nucleotides in the Blood Stages of Plasmodium falciparum

Sanz

Bandini

Ospina

et al. 2013

Journal of Biological Chemistry

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Background: GDP-fucose and other sugar nucleotide biosynthetic pathways are conserved in the P. falciparum genome. Results: These pathways are active in the intraerythrocytic life cycle of the parasite. Conclusion:The parasite biosynthesizes GDP-fucose and other sugar nucleotides not related to the glycosylphosphatidylinositol structures Significance: Their presence strongly suggests that they are involved in the biosynthesis of glycans not yet characterized.

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“…The seedlings were frozen in liquid nitrogen, homogenized with mortar and pestle, and extracted with an Isogen kit (Nippon Gene, Tokyo) according to the manufacturer's instructions. Single-strand cDNA was synthesized from approximately 2 mg of total RNA from the seedlings using a reverse-transcriptase, ReverTra Ace--(Toyobo, Osaka, Japan), and oligo(dT) [12][13][14][15][16][17][18] primers. A set of specific primers, AtUSP-F-1 (5 0 -GGA-TCCATGGCTTCTACGGTTG-3 0 ) and AtUSP-R-1 (5 0 -GAGCTCTCCATTAGAAGGACA-3 0 ), was designed based on the genomic database of Arabidopsis.…”

Section: Methodsmentioning

confidence: 99%