Fast protein affinity chromatography of two flavonoid glucosyltransferases

Latchinian, Lilian; Ekhouri, Henry E.; Ibrahim, Ragai K.

doi:10.1016/s0021-9673(01)94483-8

Cited by 18 publications

(9 citation statements)

References 9 publications

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“…Unlike other UDP-Glc-dependent glucosyltransferases (Latchinian et al, 1987;Leznicki and Bandurski, 1988;McIntosh et al, 1990), GTI did not bind strongly to either UDP-agarose or UDP-glucuronic acid agarose. When a GTI sample obtained following DEAESepharose chromatography was submitted to chromatofocusing (Polybuffer Exchanger, Pharmacia, pH 4-6) or IEF (Bio-Rad Rotofor, pH 4-6), much of the activity was lost; however, the pI of GTI was determined by chromatofocusing to be about 5.0.…”

Section: Other Chromatographic Techniques For Gtimentioning

confidence: 68%

“…The K, of GTI and GTII for UDP-Glc were 108 and 126 p~, respectively (Table 111), which is typical of UDP-Glcdependent glucosyltransferases (Shimizu and Kojima, 1984;Latchinian et al, 1987 (Table IV). In contrast to our previous report indicating that L. esculentum and L. pennellii possess similar levels of UDP-G1c:fatty acid glucosyltransferase activities (Ghangas and Steffens, 1993), under the conditions used in this study, the glucosyltransferase activity of L. esculentum was significantly higher than that of L. pennellii, particularly when the medium-chain fatty acids (8:O and 12:O) were assayed (Table IV).…”

Section: Kinetic Studiesmentioning

confidence: 96%

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Regulation of Triacylglucose Fatty Acid Composition (Uridine Diphosphate Glucose:Fatty Acid Glucosyltransferases with Overlapping Chain-Length Specificity)

1997

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UDP-glucose (UDP-ClcFoliar glandular trichomes of wild tomato (Lycopersicon pennellii) secrete a complex mixture of polar lipids composed of 2,3,4-tri-O-acylglucose esters of short-and medium-chain-length branched-and straight-chain fatty acids (Fobes et al., 1985;Burke et al., 1987). These Glc esters are responsible for the broad-spectrum insect resistance of L. pennellii (Goffreda et al., 1989; Hawthorne et al., 1992;Liedl et al., 1995). The cultivated tomato (Lycopersicon esculentum) lacks the ability to accumulate sugar esters (Fobes et al., 1985), and transfer of this trait could significantly reduce the impact of insect pests on this and other crop species (Mutschler et al., 1993 The degree to which sugar polyester biosynthesis dominates carbon flux in this species (secreted 2,3,4-tri-Oacylglucoses represent more than 20% of leaf dry weight) makes this an attractive system to study biosynthesis and metabolic control. Biosynthesis of 2,3,4-tri-O-acylglucoses in L. pennellii exhibits severa1 nove1 qualities (Fig.

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Section: Other Chromatographic Techniques For Gtimentioning

confidence: 68%

Section: Kinetic Studiesmentioning

confidence: 96%

Regulation of Triacylglucose Fatty Acid Composition (Uridine Diphosphate Glucose:Fatty Acid Glucosyltransferases with Overlapping Chain-Length Specificity)

1997

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“…1) was isolated from this plant (Bajaj et al 1983). Previous work on this enzyme indicated the close similarity, in both chromatographic (Bajaj et al 1983;Latchinian et al 1987) and kinetic (Khouri and Ibrahim 1984) properties, of the 2' -and 5 ' -0-glucosylating activities. These results suggested that O-glucosylation of these two, para-oriented positions may be catalyzed by one enzyme.…”

Section: Introductionmentioning

confidence: 61%

“…Partial purification (ca. 200-fold) of the protein pellet was performed using the Phar- macia FPLC system, by gel filtration on a column of Superose 12 (prep grade) HR 16/50, followed by ion-exchange chromatography on a column of Mono Q HR 5/5 (Latchinian et al 1987). The protein fractions which contained both 2 'and 5 ' -0-glucosylating activity were pooled and used as the source of antigen in this study.…”

Section: Extraction and Partial Purification Of Enzyme Proteinmentioning

confidence: 99%

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Characterization of a monoclonal antibody specific to a flavonol 2′-O-glucosyltransferase

Latchinian¹,

Ibrahim²

1989

Biochem. Cell Biol.

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A monoclonal antibody to a partially purified preparation of 2'-O-glucosyltransferase was produced by in vitro immunization of spleen cells from BALB/c mice, followed by fusion with mouse myeloma cells. Hybridoma culture supernatants were screened by enzyme-linked immunosorbent assay for (i) their ability to produce immunoglobulins and (ii) their immunoreactivity with a partially purified enzyme preparation. The majority of the immunoglobulin-producing hybridomas were IgM secretors. Two highly immunoreactive IgM-secreting clones were chosen for further characterization. The supernatant fraction from a culture of one of these clones displayed 50% inhibition of the 2'-O-glucosyltransferase activity. The native form of the 2'-O-glucosyltransferase was essential for recognition, suggesting that the epitope recognized by the antibody is a conformational discontiguous one.

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Immunocytochemical localization of plant secondary metabolites and the enzymes involved in their biosynthesis

Ibrahim

1990

Phytochemical Analysis

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This review highlights the basic principles involved in the immunocytochemical localization of plant secondary metabolites and their enzymes. It summarizes the methods commonly used for the preparation of antigens and characterization of antibodies, and describes the different immunocytochemical detection systems. In addition, the recent reports on immunolocalization of plant secondary metabolites and their enzymes are briefly discussed.

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Fast protein affinity chromatography of two flavonoid glucosyltransferases

Cited by 18 publications

References 9 publications

Regulation of Triacylglucose Fatty Acid Composition (Uridine Diphosphate Glucose:Fatty Acid Glucosyltransferases with Overlapping Chain-Length Specificity)

Regulation of Triacylglucose Fatty Acid Composition (Uridine Diphosphate Glucose:Fatty Acid Glucosyltransferases with Overlapping Chain-Length Specificity)

Characterization of a monoclonal antibody specific to a flavonol 2′-O-glucosyltransferase

Immunocytochemical localization of plant secondary metabolites and the enzymes involved in their biosynthesis

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