Carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts
Isolated thylakoid membranes were disrupted by treatment with nonionic detergents digitonin or dodecyl maltoside. Solubilized polypeptide complexes were separated by native gel charge shift electrophoresis. The position of ATP-synthase complex and its isolated catalytic part (CF1) within gel was det...
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National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry
2015
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ftdoajarticles:oai:doaj.org/article:c71e6aa764f04047bf0d86314f3b02ca 2023-12-03T10:21:01+01:00 Carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts A. V. Semenihin O. K. Zolotareva 2015-06-01T00:00:00Z https://doi.org/10.15407/ubj87.03.047 https://doaj.org/article/c71e6aa764f04047bf0d86314f3b02ca EN eng National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry http://ukrbiochemjournal.org/wp-content/uploads/2015/07/Semenikhin_3_15.pdf https://doaj.org/toc/2409-4943 https://doaj.org/toc/2413-5003 doi:10.15407/ubj87.03.047 2409-4943 2413-5003 https://doaj.org/article/c71e6aa764f04047bf0d86314f3b02ca The Ukrainian Biochemical Journal, Vol 87, Iss 3, Pp 47-56 (2015) ATP-synthase carbonic anhydrase coupling factor СF(1) proton transport thylakoid membranes Biochemistry QD415-436 Medicine R Biology (General) QH301-705.5 article 2015 ftdoajarticles https://doi.org/10.15407/ubj87.03.047 2023-11-05T01:38:35Z Isolated thylakoid membranes were disrupted by treatment with nonionic detergents digitonin or dodecyl maltoside. Solubilized polypeptide complexes were separated by native gel charge shift electrophoresis. The position of ATP-synthase complex and its isolated catalytic part (CF1) within gel was determined using the color reaction for ATPase activity. Due to the presence of cytochromes, the red band in unstained gels corresponded to the cytochrome b6f complex. Localization of the cytochrome b6f complex, ATP synthase and coupling CF1 in the native gel was confirmed by their subunit composition determined after SDS-electrophoretic analysis. Carbonic anhydrase (CA) activity in polypeptide zones of PS II, cytochrome b6f complex, and ATP-synthase CF1 was identified in native gels using indicator bromothymol blue. CA activity of isolated CF1 in solution was determined by infrared gas analysis as the rate of bicarbonate dehydration. The water-soluble acetazolamide, an inhibitor of CA, unlike lipophilic ethoxyzolamide inhibited CA activity of CF1. Thus, it was shown for the first time that ATP-synthase has a component which is capable of catalyzing the interconversion of forms of carbonic acid associated with proton exchange. The data obtained suggest the presence of multiple forms of carbonic anhydrase in the thylakoid membranes of spinach chloroplasts and confirm their involvement in the proton transfer to the ATP synthase. Article in Journal/Newspaper Carbonic acid Directory of Open Access Journals: DOAJ Articles The Ukrainian Biochemical Journal 87 3 47 56 |
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Directory of Open Access Journals: DOAJ Articles |
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English |
topic |
ATP-synthase carbonic anhydrase coupling factor СF(1) proton transport thylakoid membranes Biochemistry QD415-436 Medicine R Biology (General) QH301-705.5 |
spellingShingle |
ATP-synthase carbonic anhydrase coupling factor СF(1) proton transport thylakoid membranes Biochemistry QD415-436 Medicine R Biology (General) QH301-705.5 A. V. Semenihin O. K. Zolotareva Carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts |
topic_facet |
ATP-synthase carbonic anhydrase coupling factor СF(1) proton transport thylakoid membranes Biochemistry QD415-436 Medicine R Biology (General) QH301-705.5 |
description |
Isolated thylakoid membranes were disrupted by treatment with nonionic detergents digitonin or dodecyl maltoside. Solubilized polypeptide complexes were separated by native gel charge shift electrophoresis. The position of ATP-synthase complex and its isolated catalytic part (CF1) within gel was determined using the color reaction for ATPase activity. Due to the presence of cytochromes, the red band in unstained gels corresponded to the cytochrome b6f complex. Localization of the cytochrome b6f complex, ATP synthase and coupling CF1 in the native gel was confirmed by their subunit composition determined after SDS-electrophoretic analysis. Carbonic anhydrase (CA) activity in polypeptide zones of PS II, cytochrome b6f complex, and ATP-synthase CF1 was identified in native gels using indicator bromothymol blue. CA activity of isolated CF1 in solution was determined by infrared gas analysis as the rate of bicarbonate dehydration. The water-soluble acetazolamide, an inhibitor of CA, unlike lipophilic ethoxyzolamide inhibited CA activity of CF1. Thus, it was shown for the first time that ATP-synthase has a component which is capable of catalyzing the interconversion of forms of carbonic acid associated with proton exchange. The data obtained suggest the presence of multiple forms of carbonic anhydrase in the thylakoid membranes of spinach chloroplasts and confirm their involvement in the proton transfer to the ATP synthase. |
format |
Article in Journal/Newspaper |
author |
A. V. Semenihin O. K. Zolotareva |
author_facet |
A. V. Semenihin O. K. Zolotareva |
author_sort |
A. V. Semenihin |
title |
Carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts |
title_short |
Carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts |
title_full |
Carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts |
title_fullStr |
Carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts |
title_full_unstemmed |
Carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts |
title_sort |
carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts |
publisher |
National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry |
publishDate |
2015 |
url |
https://doi.org/10.15407/ubj87.03.047 https://doaj.org/article/c71e6aa764f04047bf0d86314f3b02ca |
genre |
Carbonic acid |
genre_facet |
Carbonic acid |
op_source |
The Ukrainian Biochemical Journal, Vol 87, Iss 3, Pp 47-56 (2015) |
op_relation |
http://ukrbiochemjournal.org/wp-content/uploads/2015/07/Semenikhin_3_15.pdf https://doaj.org/toc/2409-4943 https://doaj.org/toc/2413-5003 doi:10.15407/ubj87.03.047 2409-4943 2413-5003 https://doaj.org/article/c71e6aa764f04047bf0d86314f3b02ca |
op_doi |
https://doi.org/10.15407/ubj87.03.047 |
container_title |
The Ukrainian Biochemical Journal |
container_volume |
87 |
container_issue |
3 |
container_start_page |
47 |
op_container_end_page |
56 |
_version_ |
1784268581426429952 |