1-N-histidine phosphorylation of ChlD by the AAA⁺ ChlI2 stimulates magnesium chelatase activity in chlorophyll synthesis

Artur Sawicki, Zhou Shuaixiang, Kathrin Kwiatkowski, Meizhong Luo, Robert Willows

Research output: Research - peer-reviewArticle

Abstract

Magnesium chelatase (Mg-chelatase) inserts magnesium into protoporphyrin during the biosynthesis of chlorophyll and bacteriochlorophyll. Enzyme activity is reconstituted by forming two separate preactivated complexes consisting of a GUN4/ChlH/protoporphyrin IX substrate complex and a ChlI/ChlD enzyme 'motor' complex. Formation of the ChlI/ChlD complex in both Chlamydomonas reinhardtii and Oryza sativa is accompanied by phosphorylation of ChlD by ChlI, but the orthologous protein complex from Rhodobacter capsulatus, BchI/BchD, gives no detectable phosphorylation of BchD. Phosphorylation produces a 1-N-phospho-histidine within ChlD. Proteomic analysis indicates that phosphorylation occurs at a conserved His residue in the C-terminal integrin I domain of ChlD. Comparative analysis of the ChlD phosphorylation with enzyme activities of various ChlI/ChlD complexes correlates the phosphorylation by ChlI2 with stimulation of Mg-chelatase activity. Mutation of the H641 of CrChlD to E641 prevents both phosphorylation and stimulation of Mg-chelatase activity, confirming that phosphorylation at H641 stimulates Mg-chelatase. The properties of ChlI2 compared with ChlI1 of Chlamydomonas and with ChlI of Oryza, shows that ChlI2 has a regulatory role in Chlamydomonas.
LanguageEnglish
Pages2095-2105
Number of pages11
JournalBiochemical Journal
Volume474
Issue number12
DOIs
StatePublished - 2017

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Chlorophyll
Phosphorylation
magnesium chelatase
Histidine
Enzymes
Chlamydomonas
Oryza
Enzyme activity
protoporphyrin IX
Rhodobacter capsulatus
Bacteriochlorophylls
Chlamydomonas reinhardtii
Integrins
Proteomics
Mutation
Proteins
magnesium protoporphyrin
Biosynthesis
Substrates

Cite this

Sawicki, Artur ; Shuaixiang, Zhou ; Kwiatkowski, Kathrin ; Luo, Meizhong ; Willows, Robert. / 1-N-histidine phosphorylation of ChlD by the AAA⁺ ChlI2 stimulates magnesium chelatase activity in chlorophyll synthesis. In: Biochemical Journal. 2017 ; Vol. 474, No. 12. pp. 2095-2105
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abstract = "Magnesium chelatase (Mg-chelatase) inserts magnesium into protoporphyrin during the biosynthesis of chlorophyll and bacteriochlorophyll. Enzyme activity is reconstituted by forming two separate preactivated complexes consisting of a GUN4/ChlH/protoporphyrin IX substrate complex and a ChlI/ChlD enzyme 'motor' complex. Formation of the ChlI/ChlD complex in both Chlamydomonas reinhardtii and Oryza sativa is accompanied by phosphorylation of ChlD by ChlI, but the orthologous protein complex from Rhodobacter capsulatus, BchI/BchD, gives no detectable phosphorylation of BchD. Phosphorylation produces a 1-N-phospho-histidine within ChlD. Proteomic analysis indicates that phosphorylation occurs at a conserved His residue in the C-terminal integrin I domain of ChlD. Comparative analysis of the ChlD phosphorylation with enzyme activities of various ChlI/ChlD complexes correlates the phosphorylation by ChlI2 with stimulation of Mg-chelatase activity. Mutation of the H641 of CrChlD to E641 prevents both phosphorylation and stimulation of Mg-chelatase activity, confirming that phosphorylation at H641 stimulates Mg-chelatase. The properties of ChlI2 compared with ChlI1 of Chlamydomonas and with ChlI of Oryza, shows that ChlI2 has a regulatory role in Chlamydomonas.",
author = "Artur Sawicki and Zhou Shuaixiang and Kathrin Kwiatkowski and Meizhong Luo and Robert Willows",
year = "2017",
doi = "10.1042/BCJ20161094",
volume = "474",
pages = "2095--2105",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press",
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1-N-histidine phosphorylation of ChlD by the AAA⁺ ChlI2 stimulates magnesium chelatase activity in chlorophyll synthesis. / Sawicki, Artur; Shuaixiang, Zhou; Kwiatkowski, Kathrin; Luo, Meizhong; Willows, Robert.

In: Biochemical Journal, Vol. 474, No. 12, 2017, p. 2095-2105.

Research output: Research - peer-reviewArticle

TY - JOUR

T1 - 1-N-histidine phosphorylation of ChlD by the AAA⁺ ChlI2 stimulates magnesium chelatase activity in chlorophyll synthesis

AU - Sawicki,Artur

AU - Shuaixiang,Zhou

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AU - Luo,Meizhong

AU - Willows,Robert

PY - 2017

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N2 - Magnesium chelatase (Mg-chelatase) inserts magnesium into protoporphyrin during the biosynthesis of chlorophyll and bacteriochlorophyll. Enzyme activity is reconstituted by forming two separate preactivated complexes consisting of a GUN4/ChlH/protoporphyrin IX substrate complex and a ChlI/ChlD enzyme 'motor' complex. Formation of the ChlI/ChlD complex in both Chlamydomonas reinhardtii and Oryza sativa is accompanied by phosphorylation of ChlD by ChlI, but the orthologous protein complex from Rhodobacter capsulatus, BchI/BchD, gives no detectable phosphorylation of BchD. Phosphorylation produces a 1-N-phospho-histidine within ChlD. Proteomic analysis indicates that phosphorylation occurs at a conserved His residue in the C-terminal integrin I domain of ChlD. Comparative analysis of the ChlD phosphorylation with enzyme activities of various ChlI/ChlD complexes correlates the phosphorylation by ChlI2 with stimulation of Mg-chelatase activity. Mutation of the H641 of CrChlD to E641 prevents both phosphorylation and stimulation of Mg-chelatase activity, confirming that phosphorylation at H641 stimulates Mg-chelatase. The properties of ChlI2 compared with ChlI1 of Chlamydomonas and with ChlI of Oryza, shows that ChlI2 has a regulatory role in Chlamydomonas.

AB - Magnesium chelatase (Mg-chelatase) inserts magnesium into protoporphyrin during the biosynthesis of chlorophyll and bacteriochlorophyll. Enzyme activity is reconstituted by forming two separate preactivated complexes consisting of a GUN4/ChlH/protoporphyrin IX substrate complex and a ChlI/ChlD enzyme 'motor' complex. Formation of the ChlI/ChlD complex in both Chlamydomonas reinhardtii and Oryza sativa is accompanied by phosphorylation of ChlD by ChlI, but the orthologous protein complex from Rhodobacter capsulatus, BchI/BchD, gives no detectable phosphorylation of BchD. Phosphorylation produces a 1-N-phospho-histidine within ChlD. Proteomic analysis indicates that phosphorylation occurs at a conserved His residue in the C-terminal integrin I domain of ChlD. Comparative analysis of the ChlD phosphorylation with enzyme activities of various ChlI/ChlD complexes correlates the phosphorylation by ChlI2 with stimulation of Mg-chelatase activity. Mutation of the H641 of CrChlD to E641 prevents both phosphorylation and stimulation of Mg-chelatase activity, confirming that phosphorylation at H641 stimulates Mg-chelatase. The properties of ChlI2 compared with ChlI1 of Chlamydomonas and with ChlI of Oryza, shows that ChlI2 has a regulatory role in Chlamydomonas.

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