title: The light-harvesting chlorophyll a/b complex can be reconstituted in vitro from its completely unfolded apoprotein.
abstract: The major light-harvesting chlorophyll a/b protein (LHCIIb) of higher plants is one of the few membrane proteins that can be refolded in vitro. During folding, the apoprotein is assembled with pigments to form a structurally authentic and functional pigment--protein complex. All reconstitution procedures used so far include solubilization of the apoprotein in sodium dodecyl sulfate (SDS) where the protein adopts approximately half of its alpha-helical folding present in the native structure. This paper shows that this preformed alpha-helix is not a prerequisite for LHCIIb folding in vitro. The apoprotein can also be reconstituted starting from a solution in guanidinium hydrochloride (Gnd) where the protein contains no detectable helical structure. Reconstitution yields are somewhat lower in the Gnd than in the SDS procedure, but the reconstitution products exhibit very similar biochemical and spectroscopic properties. The kinetics of LHCIIb assembly, as assessed by time-resolved fluorescence measurements, are virtually the same in both reconstitution procedures. This demonstrates that the initiation of alpha-helix formation is not a rate-limiting step in LHCIIb apoprotein folding.
authors.affiliation: Institut für Allgemeine Botanik der Johannes-Gutenberg-Universität, Müllerweg 6, D-55099 Mainz, Germany.
authors.forename: Chunhong
authors.initials: C
authors.lastname: Yang
authors.forename: Ruth
authors.initials: R
authors.lastname: Horn
authors.forename: Harald
authors.initials: H
authors.lastname: Paulsen
mesh_terms: D001059:Apoproteins / Q000378:metabolism*; D002734:Chlorophyll / Q000378:metabolism; D000077194:Chlorophyll A; D002942:Circular Dichroism; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism*; D017510:Protein Folding; D013050:Spectrometry, Fluorescence
chemical_list: D001059:Apoproteins; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D002734:Chlorophyll; C037184:chlorophyll b; D000077194:Chlorophyll A