Substrate-binding model of the chlorophyll biosynthetic magnesium chelatase BchH subunit.
(2008) In Journal of Biological Chemistry 283(17). p.11652-11660- Abstract
- Photosynthetic organisms require chlorophyll and bacteriochlorophyll to harness light energy and transform water and carbon dioxide into carbohydrates and oxygen. The biosynthesis of these pigments is initiated by magnesium chelatase, an enzyme composed of BchI, BchD and BchH proteins, which catalyzes the insertion of Mg(2+) into protoporphyrin IX (Proto) to produce Mg-protoporphyrin IX. BchI and BchD form an ATP dependant AAA(+) complex that transiently interacts with the Proto-binding BchH subunit, at which point Mg(2+) is chelated. In this study, controlled proteolysis, electron microscopy of negatively stained specimens and single-particle 3D reconstruction have been used to probe the structure and substrate binding mechanism of the... (More)
- Photosynthetic organisms require chlorophyll and bacteriochlorophyll to harness light energy and transform water and carbon dioxide into carbohydrates and oxygen. The biosynthesis of these pigments is initiated by magnesium chelatase, an enzyme composed of BchI, BchD and BchH proteins, which catalyzes the insertion of Mg(2+) into protoporphyrin IX (Proto) to produce Mg-protoporphyrin IX. BchI and BchD form an ATP dependant AAA(+) complex that transiently interacts with the Proto-binding BchH subunit, at which point Mg(2+) is chelated. In this study, controlled proteolysis, electron microscopy of negatively stained specimens and single-particle 3D reconstruction have been used to probe the structure and substrate binding mechanism of the BchH subunit to a resolution of 25 A. The apo structure contains three major lobe-shaped domains connected at a single point, with additional densities at the tip of two lobes termed the thumb and finger. With the independent reconstruction of a substrate bound BchH complex (BchH-Proto) we observed a distinct conformational change in the thumb and finger subdomains. Prolonged proteolysis of native apo BchH produced a stable C-terminal fragment of 45 kDa, and Proto was shown to protect the full polypeptide from degradation. Fitting of a truncated BchH polypeptide reconstruction identified the N- and C-terminal domains. Our results show that the N- and C-terminal domains play crucial roles in the substrate-binding mechanism. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1042097
- author
- Sirijovski, Nickolche LU ; Lundqvist, Joakim LU ; Rosenbäck, Matilda ; Elmlund, Hans LU ; Al-Karadaghi, Salam LU ; Willows, Robert D and Hansson, Mats LU
- organization
- publishing date
- 2008
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 283
- issue
- 17
- pages
- 11652 - 11660
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- pmid:18263581
- wos:000255067400066
- scopus:45549091468
- pmid:18263581
- ISSN
- 1083-351X
- DOI
- 10.1074/jbc.M709172200
- language
- English
- LU publication?
- yes
- additional info
- Papers In Press, published online ahead of print February 8, 2008
- id
- e7ed5463-161c-4b7d-9b64-f4d320b79e3b (old id 1042097)
- date added to LUP
- 2016-04-01 12:02:28
- date last changed
- 2022-03-13 04:28:19
@article{e7ed5463-161c-4b7d-9b64-f4d320b79e3b, abstract = {{Photosynthetic organisms require chlorophyll and bacteriochlorophyll to harness light energy and transform water and carbon dioxide into carbohydrates and oxygen. The biosynthesis of these pigments is initiated by magnesium chelatase, an enzyme composed of BchI, BchD and BchH proteins, which catalyzes the insertion of Mg(2+) into protoporphyrin IX (Proto) to produce Mg-protoporphyrin IX. BchI and BchD form an ATP dependant AAA(+) complex that transiently interacts with the Proto-binding BchH subunit, at which point Mg(2+) is chelated. In this study, controlled proteolysis, electron microscopy of negatively stained specimens and single-particle 3D reconstruction have been used to probe the structure and substrate binding mechanism of the BchH subunit to a resolution of 25 A. The apo structure contains three major lobe-shaped domains connected at a single point, with additional densities at the tip of two lobes termed the thumb and finger. With the independent reconstruction of a substrate bound BchH complex (BchH-Proto) we observed a distinct conformational change in the thumb and finger subdomains. Prolonged proteolysis of native apo BchH produced a stable C-terminal fragment of 45 kDa, and Proto was shown to protect the full polypeptide from degradation. Fitting of a truncated BchH polypeptide reconstruction identified the N- and C-terminal domains. Our results show that the N- and C-terminal domains play crucial roles in the substrate-binding mechanism.}}, author = {{Sirijovski, Nickolche and Lundqvist, Joakim and Rosenbäck, Matilda and Elmlund, Hans and Al-Karadaghi, Salam and Willows, Robert D and Hansson, Mats}}, issn = {{1083-351X}}, language = {{eng}}, number = {{17}}, pages = {{11652--11660}}, publisher = {{American Society for Biochemistry and Molecular Biology}}, series = {{Journal of Biological Chemistry}}, title = {{Substrate-binding model of the chlorophyll biosynthetic magnesium chelatase BchH subunit.}}, url = {{http://dx.doi.org/10.1074/jbc.M709172200}}, doi = {{10.1074/jbc.M709172200}}, volume = {{283}}, year = {{2008}}, }