Isolation of the rotenome-sensitive NADH-ubiquinone reductase (complex I) from red beet mitochondria
(1994) In Physiologia Plantarum 90(3). p.607-615- Abstract
- Complex 1 of the respirator) chain (EC 1.6.531, measured as NADH-duroquinone and NADH-ubiquinone, reductase activities, was isolated from purified red beetroot (Beta vulgaris L.I mitochondria. The mitochondria were disrupted by freeze-thawing and inner membrane vesicles were pelleted. After solubilization of the vesicles with Triton X-100, the enzyme complex was purified 11-fold (compared to the activity in the inner membrane vesicles) by size-exclusion chromatography on a Sephacryl S-400 HR column and then by ion-exchange chromatography on a DEAE-Sepharose CL-6B column. Triton X-100 was present throughout the purification procedure. Tire purified complex showed approximately 30 bands on SDS-PAGE and about 15 polypeptides including those... (More)
- Complex 1 of the respirator) chain (EC 1.6.531, measured as NADH-duroquinone and NADH-ubiquinone, reductase activities, was isolated from purified red beetroot (Beta vulgaris L.I mitochondria. The mitochondria were disrupted by freeze-thawing and inner membrane vesicles were pelleted. After solubilization of the vesicles with Triton X-100, the enzyme complex was purified 11-fold (compared to the activity in the inner membrane vesicles) by size-exclusion chromatography on a Sephacryl S-400 HR column and then by ion-exchange chromatography on a DEAE-Sepharose CL-6B column. Triton X-100 was present throughout the purification procedure. Tire purified complex showed approximately 30 bands on SDS-PAGE and about 15 polypeptides including those at 80. 54, 53. 51. 27. 25 and 22 kDa cross-reacted with polyclonal antibodies raised against complex I from Neurospora crassa. This is similar lo the pattern obtained with complex I from Neurospera crassa.
Analysis by nativc-SDS 2-dimensional PAGE revealed the existence of several molecular mass forms of the purified complex.
After reconstitution of the purified complex into phosphatidylcholine vesicles, the NADH-ubiquinone reductase activity had a Km (NADH) of about I μM and was inhibited by both rotenone and dicyclohexylcarbodiimide.
(Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/3407dfee-7c7d-4c09-8159-4dcc30069206
- author
- Rasmusson, A. G. LU ; Mendel-Hartvig, J. ; Moller, Ian M. and Wiskich, J. T.
- organization
- publishing date
- 1994
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Beta vulgaris, complex I, inner membrane, mitochondrion, NADH dehydrogenase, red beetroot
- in
- Physiologia Plantarum
- volume
- 90
- issue
- 3
- pages
- 607 - 615
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- scopus:0028025890
- ISSN
- 0031-9317
- DOI
- 10.1034/j.1399-3054.1994.900324.x
- language
- English
- LU publication?
- yes
- id
- 3407dfee-7c7d-4c09-8159-4dcc30069206
- date added to LUP
- 2016-05-31 21:37:40
- date last changed
- 2024-01-04 07:46:26
@article{3407dfee-7c7d-4c09-8159-4dcc30069206, abstract = {{Complex 1 of the respirator) chain (EC 1.6.531, measured as NADH-duroquinone and NADH-ubiquinone, reductase activities, was isolated from purified red beetroot (Beta vulgaris L.I mitochondria. The mitochondria were disrupted by freeze-thawing and inner membrane vesicles were pelleted. After solubilization of the vesicles with Triton X-100, the enzyme complex was purified 11-fold (compared to the activity in the inner membrane vesicles) by size-exclusion chromatography on a Sephacryl S-400 HR column and then by ion-exchange chromatography on a DEAE-Sepharose CL-6B column. Triton X-100 was present throughout the purification procedure. Tire purified complex showed approximately 30 bands on SDS-PAGE and about 15 polypeptides including those at 80. 54, 53. 51. 27. 25 and 22 kDa cross-reacted with polyclonal antibodies raised against complex I from Neurospora crassa. This is similar lo the pattern obtained with complex I from Neurospera crassa.<br/>Analysis by nativc-SDS 2-dimensional PAGE revealed the existence of several molecular mass forms of the purified complex.<br/>After reconstitution of the purified complex into phosphatidylcholine vesicles, the NADH-ubiquinone reductase activity had a Km (NADH) of about I μM and was inhibited by both rotenone and dicyclohexylcarbodiimide.<br/>}}, author = {{Rasmusson, A. G. and Mendel-Hartvig, J. and Moller, Ian M. and Wiskich, J. T.}}, issn = {{0031-9317}}, keywords = {{Beta vulgaris; complex I; inner membrane; mitochondrion; NADH dehydrogenase; red beetroot}}, language = {{eng}}, number = {{3}}, pages = {{607--615}}, publisher = {{John Wiley & Sons Inc.}}, series = {{Physiologia Plantarum}}, title = {{Isolation of the rotenome-sensitive NADH-ubiquinone reductase (complex I) from red beet mitochondria}}, url = {{http://dx.doi.org/10.1034/j.1399-3054.1994.900324.x}}, doi = {{10.1034/j.1399-3054.1994.900324.x}}, volume = {{90}}, year = {{1994}}, }