Role of the C-Terminal Extrinsic Region of the α Polypeptide of the Light-Harvesting 2 Complex of Rhodobacter sphaeroides: A Domain Swap Study
| العنوان: | Role of the C-Terminal Extrinsic Region of the α Polypeptide of the Light-Harvesting 2 Complex of Rhodobacter sphaeroides: A Domain Swap Study |
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| المؤلفون: | John D. Olsen, C. Alistair Siebert, C. Neil Hunter, Per A. Bullough, Bruno Robert |
| المصدر: | Biochemistry. 42:15114-15123 |
| بيانات النشر: | American Chemical Society (ACS), 2003. |
| سنة النشر: | 2003 |
| مصطلحات موضوعية: | Circular dichroism, Hot Temperature, Recombinant Fusion Proteins, Molecular Sequence Data, Mutant, Light-Harvesting Protein Complexes, Rhodobacter sphaeroides, Spectrum Analysis, Raman, Ring (chemistry), Biochemistry, chemistry.chemical_compound, Protein structure, Bacterial Proteins, Amino Acid Sequence, Rhodospirillum, Sequence Deletion, biology, Circular Dichroism, biology.organism_classification, Carotenoids, Peptide Fragments, Protein Structure, Tertiary, Ring size, Protein Subunits, Crystallography, Spectrometry, Fluorescence, chemistry, Spectrophotometry, Bacteriochlorophyll, Crystallization, Neurosporene |
| الوصف: | The LH1 and LH2 complexes of Rhodobacter sphaeroides form ring structures of 16 and 9 protomers, respectively, comprising alpha and beta polypeptides, bacteriochlorophylls (Bchl), and carotenoids. Using the LH2 complex as a starting point, two chimeric LH complexes were constructed incorporating the alphaC-terminal domain of either the Rb. sphaeroides LH1 complex or the Rhodospirillum molischianum LH2 complex. The LH1 domain swap produced a new red-shifted component that comprised approximately 30% of the total absorbance. In the LH1alpha C-terminal mutant this new red-shifted species acts as the terminal emitter, with the new emission maximum located 10 nm further to the red than for the WT. Raman spectroscopy indicates that a fraction of the B850 Bchls is involved in relatively weak H-bonds, possibly involving the alphaTrp(+11) residue within the new alphaC-terminus, consistent with a more LH1-like character for one of the Bchls. The CD data indicate that the domain swaps have perturbed the native arrangement of the B850 Bchls, including the site energy difference between the alpha- and beta-bound Bchls. Thus, the normal energetic structure of the ring system has been disrupted, with one component blue shifted due to the presumed loss of an H-bond donor and the other red shifted by the influence of the new alphaC-terminal domain. The dichotomous response of the mutants to the carotenoids incorporated, spheroidenone or neurosporene, strongly suggests that the C-terminal region of the alpha polypeptide is involved in binding a carotenoid. The projection map of the LH1alpha C-terminal mutant complex was determined in negative stain at 25 A resolution, and it shows a diameter of 53 A, compared to 50 A for the WT. Hence these new spectral properties have not been accompanied by an alteration in ring size. |
| تدمد: | 1520-4995 0006-2960 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9a1e8fc43e741da0c8b3d249cc3c373eTest https://doi.org/10.1021/bi035411hTest |
| رقم الانضمام: | edsair.doi.dedup.....9a1e8fc43e741da0c8b3d249cc3c373e |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15204995 00062960 |
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