Isolation and polypeptide composition of 1,3-ß-glucan synthase from plasma membranes of Brassica oleracea
(1991) In Physiologia Plantarum 81(3). p.289-294- Abstract
- The l,3‐ß‐glucan synthase (callose synthase, EC 2.4.1.34) was solubilized from cauliflower (Brassica oleracea L.) plasma membranes with digitonin, and partially purified by ion exchange chromatography and gel filtration [fast protein liquid chromatography (FPLC)] using 3‐[(cholamidopropyl)dimethylammonio]‐1‐propane‐sulfonate (CHAPS) in the elution buffers. These initial steps were necessary to obtain specific precipitation of the enzyme during product entrapment, the final purification step. Five polypeptides of 32, 35, 57, 65 and 66 kDa were highly enriched in the final preparation and are thus likely components of the callose synthase complex. The purified enzyme was activated by Ca2+, spermine and cellobiose in the same way... (More)
- The l,3‐ß‐glucan synthase (callose synthase, EC 2.4.1.34) was solubilized from cauliflower (Brassica oleracea L.) plasma membranes with digitonin, and partially purified by ion exchange chromatography and gel filtration [fast protein liquid chromatography (FPLC)] using 3‐[(cholamidopropyl)dimethylammonio]‐1‐propane‐sulfonate (CHAPS) in the elution buffers. These initial steps were necessary to obtain specific precipitation of the enzyme during product entrapment, the final purification step. Five polypeptides of 32, 35, 57, 65 and 66 kDa were highly enriched in the final preparation and are thus likely components of the callose synthase complex. The purified enzyme was activated by Ca2+, spermine and cellobiose in the same way as the enzyme in situ, indicating that no essential subunits were missing. The polyglucan produced by the purified enzyme contained mainly 1,3‐linked glucose. (Less)
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https://lup.lub.lu.se/record/0560132a-759c-4be7-bc09-877e9cafce6d
- author
- Fredrikson, Karin ; Kjellbom, Per LU and Larsson, Christer LU
- organization
- publishing date
- 1991
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Brassica oleracea, callose synthase, cauliflower, 1,3‐ß‐glucan synthase, plasma membranes, polypeptide composition, protein purification
- in
- Physiologia Plantarum
- volume
- 81
- issue
- 3
- pages
- 5 pages
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- scopus:0043033349
- ISSN
- 0031-9317
- DOI
- 10.1111/j.1399-3054.1991.tb08734.x
- language
- English
- LU publication?
- yes
- id
- 0560132a-759c-4be7-bc09-877e9cafce6d
- date added to LUP
- 2019-06-20 17:00:42
- date last changed
- 2021-03-21 04:09:03
@article{0560132a-759c-4be7-bc09-877e9cafce6d, abstract = {{The l,3‐ß‐glucan synthase (callose synthase, EC 2.4.1.34) was solubilized from cauliflower (Brassica oleracea L.) plasma membranes with digitonin, and partially purified by ion exchange chromatography and gel filtration [fast protein liquid chromatography (FPLC)] using 3‐[(cholamidopropyl)dimethylammonio]‐1‐propane‐sulfonate (CHAPS) in the elution buffers. These initial steps were necessary to obtain specific precipitation of the enzyme during product entrapment, the final purification step. Five polypeptides of 32, 35, 57, 65 and 66 kDa were highly enriched in the final preparation and are thus likely components of the callose synthase complex. The purified enzyme was activated by Ca<sup>2+</sup>, spermine and cellobiose in the same way as the enzyme in situ, indicating that no essential subunits were missing. The polyglucan produced by the purified enzyme contained mainly 1,3‐linked glucose.}}, author = {{Fredrikson, Karin and Kjellbom, Per and Larsson, Christer}}, issn = {{0031-9317}}, keywords = {{Brassica oleracea; callose synthase; cauliflower; 1,3‐ß‐glucan synthase; plasma membranes; polypeptide composition; protein purification}}, language = {{eng}}, number = {{3}}, pages = {{289--294}}, publisher = {{John Wiley & Sons Inc.}}, series = {{Physiologia Plantarum}}, title = {{Isolation and polypeptide composition of 1,3-ß-glucan synthase from plasma membranes of Brassica oleracea}}, url = {{http://dx.doi.org/10.1111/j.1399-3054.1991.tb08734.x}}, doi = {{10.1111/j.1399-3054.1991.tb08734.x}}, volume = {{81}}, year = {{1991}}, }