Skip to Main Content
Assays for the activities of polyamine biosynthetic enzymes using intact tissuesAuthor(s): Rakesh Minocha; Stephanie Long; Hisae Maki; Subhash C. Minocha
Source: Plant Physiology and Biochemistry. 37: 597-603.
Publication Series: Scientific Journal (JRNL)
Station: Northern Research Station
PDF: Download Publication (729.97 KB)
DescriptionTraditionally, most enzyme assays utilize homogenized cell extracts with or without dialysis. Homogenization and centrifugation of large numbers of samples for screening of mutants and transgenic cell lines is quite cumbersome and generally requires sufficiently large amounts (hundreds of milligrams) of tissue. However, in situations where the tissue is available in small quantities, or one needs to study changes in enzyme activities during development (e.g. somatic embryogenesis), it is desirable to have rapid and reproducible assay methods that utilize only a few milligrams of tissue and can be conducted without homogenization. Here, we report a procedure for the measurement of enzyme activities of the three key decarboxylases involved in polyamine biosynthesis utilizing small quantities of plant tissue without the homogenization and centrifugation steps. Suspension cultures of red spruce (Picea rubens (Sarg.)), hybrid poplar (Populus nigra x maximowiczii), and wild carrot (Daucus carota) were used directly to measure decarboxylation of omithine, arginine and S-adenosylmethionine. Our results demonstrate that this procedure can be used to quantify the activities of arginine decarboxylase (EC 184.108.40.206), omithine decarboxylase (EC 220.127.116.11) and S-adenosylmethionine decarboxylase (EC 4.1.l.50) in a manner quite comparable to the traditional assays for these enzymes that involve laborious steps of homogenization and centrifugation.
- Check the Northern Research Station web site to request a printed copy of this publication.
- Our on-line publications are scanned and captured using Adobe Acrobat.
- During the capture process some typographical errors may occur.
- Please contact Sharon Hobrla, firstname.lastname@example.org if you notice any errors which make this publication unusable.
- We recommend that you also print this page and attach it to the printout of the article, to retain the full citation information.
- This article was written and prepared by U.S. Government employees on official time, and is therefore in the public domain.
CitationMinocha, Rakesh; Long, Stephanie; Maki, Hisae; Minocha, Subhash C. 1999. Assays for the activities of polyamine biosynthetic enzymes using intact tissues. Plant Physiology and Biochemistry. 37: 597-603.
KeywordsADC, carrot, enzyme assays, ODC, polyamines, SAMDC, trees
- The effects of genetic manipulation of putrescine biosynthesis on transcription and activities of the other polyamine biosynthetic enzymes
- Transgenic manipulation of a single polyamine in poplar cells affects the accumulation of all amino acids
- Transgenic manipulation of the metabolism of polyamines in poplar cells
XML: View XML