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Isolation and polypeptide composition of 1,3-ß-glucan synthase from plasma membranes of Brassica oleracea

Fredrikson, Karin ; Kjellbom, Per LU and Larsson, Christer LU (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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author
; and
organization
publishing date
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}},
}