Functionally important domains of the large hydrophilic loop of CP47 as probed by oligonucleotide-directed mutagenesis in Synechocystis sp. PCC 6803

Elisabeth Haag, Julian J. Eaton-Rye, Gernot Renger, Willem Vermaas

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

The chlorophyll a-binding protein CP47 serves as core antenna to photosystem II (PS II). The predicted topology of CP47 exhibits six membrane-spanning regions and a large hydrophilic loop (loop E) which rougly includes 200 residues (255-455) and is presumably exposed to the lumenal side of the thylakoid membrane. Several lines of experimental evidence suggest that loop E might be involved in binding or stabilizing functional manganese in the catalytic site of water oxidation or in interacting with the extrinsic PS II-O protein (the 33-kDa manganese-stabilizing protein). To scan loop E for functionally important domains, oligonucleotide-directed mutagenesis has been used to introduce deletions of 3-8 residues in conserved and charged regions of loop E. In addition, one single-site mutation of the only histidine present in loop E was created (H343L). Domains deleted in Δ1 (I265-F268), Δ2 (T271-K277), Δ4 (T304-L309), Δ5 (F311-N317), and Δ12 (D440-P447) are required for stable assembly of functional PS II complexes. Deletion of domains Δ3 (K277-E283) and Δ11 (R422-E428) significantly reduces the level of assembled PS II and impairs photoautotrophic growth and oxygen evolution. Deletion of domain Δ8 (A373-D380) enhances the susceptibility to photoinhibition. In contrast, deletion of domains Δ6 (G333-I336), Δ7 (K347-R352), Δ9 (V392-Q394), and Δ10 (D416-F420) and mutation of H343 to leucine do not seem to severly interrupt PS II structure and function, although all mutants exhibit a slightly decreased stability of PS II as compared to the wild type. Thus, selected domains of the large hydrophilic loop of CP47 are important for PS II structure and function. With respect to possible sites of interaction between loop E of CP47 and the extrinsic PS II-O protein, our results indicate that none of the deletions in the region from residue 330 to 420 (Δ6, Δ7, Δ8, Δ9, Δ0) completely interrupts a functional association of the manganese-stabilizing protein to PS II, although the binding characteristics might be changed in some cases.

Original languageEnglish (US)
Pages (from-to)4444-4454
Number of pages11
JournalBiochemistry
Volume32
Issue number16
StatePublished - 1993

Fingerprint

Synechocystis
Mutagenesis
Photosystem II Protein Complex
Site-Directed Mutagenesis
Oligonucleotides
Manganese
Proteins
Mutation
Thylakoids
Membranes
Histidine
Leucine
Catalytic Domain
Oxygen
Topology
Antennas
Water
Oxidation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Functionally important domains of the large hydrophilic loop of CP47 as probed by oligonucleotide-directed mutagenesis in Synechocystis sp. PCC 6803. / Haag, Elisabeth; Eaton-Rye, Julian J.; Renger, Gernot; Vermaas, Willem.

In: Biochemistry, Vol. 32, No. 16, 1993, p. 4444-4454.

Research output: Contribution to journalArticle

@article{ae883a7109784f939a11f681eb932e4d,
title = "Functionally important domains of the large hydrophilic loop of CP47 as probed by oligonucleotide-directed mutagenesis in Synechocystis sp. PCC 6803",
abstract = "The chlorophyll a-binding protein CP47 serves as core antenna to photosystem II (PS II). The predicted topology of CP47 exhibits six membrane-spanning regions and a large hydrophilic loop (loop E) which rougly includes 200 residues (255-455) and is presumably exposed to the lumenal side of the thylakoid membrane. Several lines of experimental evidence suggest that loop E might be involved in binding or stabilizing functional manganese in the catalytic site of water oxidation or in interacting with the extrinsic PS II-O protein (the 33-kDa manganese-stabilizing protein). To scan loop E for functionally important domains, oligonucleotide-directed mutagenesis has been used to introduce deletions of 3-8 residues in conserved and charged regions of loop E. In addition, one single-site mutation of the only histidine present in loop E was created (H343L). Domains deleted in Δ1 (I265-F268), Δ2 (T271-K277), Δ4 (T304-L309), Δ5 (F311-N317), and Δ12 (D440-P447) are required for stable assembly of functional PS II complexes. Deletion of domains Δ3 (K277-E283) and Δ11 (R422-E428) significantly reduces the level of assembled PS II and impairs photoautotrophic growth and oxygen evolution. Deletion of domain Δ8 (A373-D380) enhances the susceptibility to photoinhibition. In contrast, deletion of domains Δ6 (G333-I336), Δ7 (K347-R352), Δ9 (V392-Q394), and Δ10 (D416-F420) and mutation of H343 to leucine do not seem to severly interrupt PS II structure and function, although all mutants exhibit a slightly decreased stability of PS II as compared to the wild type. Thus, selected domains of the large hydrophilic loop of CP47 are important for PS II structure and function. With respect to possible sites of interaction between loop E of CP47 and the extrinsic PS II-O protein, our results indicate that none of the deletions in the region from residue 330 to 420 (Δ6, Δ7, Δ8, Δ9, Δ0) completely interrupts a functional association of the manganese-stabilizing protein to PS II, although the binding characteristics might be changed in some cases.",
author = "Elisabeth Haag and Eaton-Rye, {Julian J.} and Gernot Renger and Willem Vermaas",
year = "1993",
language = "English (US)",
volume = "32",
pages = "4444--4454",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "16",

}

TY - JOUR

T1 - Functionally important domains of the large hydrophilic loop of CP47 as probed by oligonucleotide-directed mutagenesis in Synechocystis sp. PCC 6803

AU - Haag, Elisabeth

AU - Eaton-Rye, Julian J.

AU - Renger, Gernot

AU - Vermaas, Willem

PY - 1993

Y1 - 1993

N2 - The chlorophyll a-binding protein CP47 serves as core antenna to photosystem II (PS II). The predicted topology of CP47 exhibits six membrane-spanning regions and a large hydrophilic loop (loop E) which rougly includes 200 residues (255-455) and is presumably exposed to the lumenal side of the thylakoid membrane. Several lines of experimental evidence suggest that loop E might be involved in binding or stabilizing functional manganese in the catalytic site of water oxidation or in interacting with the extrinsic PS II-O protein (the 33-kDa manganese-stabilizing protein). To scan loop E for functionally important domains, oligonucleotide-directed mutagenesis has been used to introduce deletions of 3-8 residues in conserved and charged regions of loop E. In addition, one single-site mutation of the only histidine present in loop E was created (H343L). Domains deleted in Δ1 (I265-F268), Δ2 (T271-K277), Δ4 (T304-L309), Δ5 (F311-N317), and Δ12 (D440-P447) are required for stable assembly of functional PS II complexes. Deletion of domains Δ3 (K277-E283) and Δ11 (R422-E428) significantly reduces the level of assembled PS II and impairs photoautotrophic growth and oxygen evolution. Deletion of domain Δ8 (A373-D380) enhances the susceptibility to photoinhibition. In contrast, deletion of domains Δ6 (G333-I336), Δ7 (K347-R352), Δ9 (V392-Q394), and Δ10 (D416-F420) and mutation of H343 to leucine do not seem to severly interrupt PS II structure and function, although all mutants exhibit a slightly decreased stability of PS II as compared to the wild type. Thus, selected domains of the large hydrophilic loop of CP47 are important for PS II structure and function. With respect to possible sites of interaction between loop E of CP47 and the extrinsic PS II-O protein, our results indicate that none of the deletions in the region from residue 330 to 420 (Δ6, Δ7, Δ8, Δ9, Δ0) completely interrupts a functional association of the manganese-stabilizing protein to PS II, although the binding characteristics might be changed in some cases.

AB - The chlorophyll a-binding protein CP47 serves as core antenna to photosystem II (PS II). The predicted topology of CP47 exhibits six membrane-spanning regions and a large hydrophilic loop (loop E) which rougly includes 200 residues (255-455) and is presumably exposed to the lumenal side of the thylakoid membrane. Several lines of experimental evidence suggest that loop E might be involved in binding or stabilizing functional manganese in the catalytic site of water oxidation or in interacting with the extrinsic PS II-O protein (the 33-kDa manganese-stabilizing protein). To scan loop E for functionally important domains, oligonucleotide-directed mutagenesis has been used to introduce deletions of 3-8 residues in conserved and charged regions of loop E. In addition, one single-site mutation of the only histidine present in loop E was created (H343L). Domains deleted in Δ1 (I265-F268), Δ2 (T271-K277), Δ4 (T304-L309), Δ5 (F311-N317), and Δ12 (D440-P447) are required for stable assembly of functional PS II complexes. Deletion of domains Δ3 (K277-E283) and Δ11 (R422-E428) significantly reduces the level of assembled PS II and impairs photoautotrophic growth and oxygen evolution. Deletion of domain Δ8 (A373-D380) enhances the susceptibility to photoinhibition. In contrast, deletion of domains Δ6 (G333-I336), Δ7 (K347-R352), Δ9 (V392-Q394), and Δ10 (D416-F420) and mutation of H343 to leucine do not seem to severly interrupt PS II structure and function, although all mutants exhibit a slightly decreased stability of PS II as compared to the wild type. Thus, selected domains of the large hydrophilic loop of CP47 are important for PS II structure and function. With respect to possible sites of interaction between loop E of CP47 and the extrinsic PS II-O protein, our results indicate that none of the deletions in the region from residue 330 to 420 (Δ6, Δ7, Δ8, Δ9, Δ0) completely interrupts a functional association of the manganese-stabilizing protein to PS II, although the binding characteristics might be changed in some cases.

UR - http://www.scopus.com/inward/record.url?scp=0027299939&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027299939&partnerID=8YFLogxK

M3 - Article

VL - 32

SP - 4444

EP - 4454

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 16

ER -