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Differential assembly of polypeptides of the light-harvesting 2 complex encoded by distinct operons during acclimation of Rhodobacter sphaeroides to low light intensity

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Abstract

In order to obtain an improved understanding of the assembly of the bacterial photosynthetic apparatus, we have conducted a proteomic analysis of pigment-protein complexes isolated from the purple bacterium Rhodobacter sphaeroides undergoing acclimation to reduced incident light intensity. Photoheterotrophically growing cells were shifted from 1,100 to 100 W/m2 and intracytoplasmic membrane (ICM) vesicles isolated over 24-h were subjected to clear native polyacrylamide gel electrophoresis. Bands containing the LH2 and reaction center (RC)-LH1 complexes were excised and subjected to in-gel trypsin digestion followed by liquid chromatography (LC)-mass spectroscopy (MS)/MS. The results revealed that the LH2 band contained distinct levels of the LH2-α and -β polypeptides encoded by the two puc operons. Polypeptide subunits encoded by the puc2AB operon predominated under high light and in the early stages of acclimation to low light, while after 24 h, the puc1BAC components were most abundant. Surprisingly, the Puc2A polypeptide containing a 251 residue C-terminal extension not present in Puc1A, was a protein of major abundance. A predominance of Puc2A components in the LH2 complex formed at high light intensity is followed by a >2.5-fold enrichment in Puc1B levels between 3 and 24 h of acclimation, accompanied by a nearly twofold decrease in Puc2A levels. This indicates that the puc1BAC operon is under more stringent light control, thought to reflect differences in the puc1 upstream regulatory region. In contrast, elevated levels of Puc2 polypeptides were seen 48 h after the gratuitous induction of ICM formation at low aeration in the dark, while after 24 h of acclimation to low light, an absence of alterations in Puc polypeptide distributions was observed in the upper LH2-enriched gel band, despite an approximate twofold increase in overall LH2 levels. This is consistent with the origin of this band from a pool of LH2 laid down early in development that is distinct from subsequently assembled LH2-only domains, forming the LH2 gel band.

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Abbreviations

AFM:

Atomic force microscopy

β-OG:

n-octyl β-d-glucopyranoside

DOC:

Deoxycholate

CM:

Cytoplasmic membrane

COGs:

Clusters of orthologous groups

ICM:

Intracytoplasmic membrane

LH:

Light harvesting

LH1:

Core pigment-protein light-harvesting complex

LH2:

Peripheral pigment-protein light-harvesting complex

RC:

Photochemical reaction center

UPB:

Upper pigmented band

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Acknowledgments

We thank Prof. Peter Lobel and Dr. Haiyan Zheng of the Center for Advanced Biotechnology and Medicine, University of Medicine and Dentistry of New Jersey, for conducting the proteomics analysis. This work was supported by the Aresty Research Center for Undergraduates at Rutgers University (OBO, HCS, JTL) and U. S. Department of Energy Grant No. DE-FG02-08ER15957 from the Chemical Sciences, Geosciences and Biosciences Division, Office of Basic Energy Sciences, Office of Science (RAN).

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  1. Department of Molecular Biology and Biochemistry, Rutgers University, Busch Campus, 604 Allison Road, Piscataway, NJ, 08854-8082, USA

    Kamil Woronowicz, Oluwatobi B. Olubanjo, Hee Chang Sung, Joana L. Lamptey & Robert A. Niederman

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  1. Kamil Woronowicz
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  2. Oluwatobi B. Olubanjo
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  3. Hee Chang Sung
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  4. Joana L. Lamptey
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  5. Robert A. Niederman
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Correspondence to Robert A. Niederman.

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This is a re-publication of the original article in the September 2011 issue of this journal. When referring to this article in future publications, please cite the original article, as follows: Woronowicz K, Olubanjo OB, Sung HC, Lamptey JL, Niederman RA (2011) Differential assembly of polypeptides of the light-harvesting 2 complex encoded by distinct operons during acclimation of Rhodobacter sphaeroides to low light intensity. Photosynth Res 108(2–3):201–214. doi: 10.1007/s11120-011-9681-x

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Woronowicz, K., Olubanjo, O.B., Sung, H.C. et al. Differential assembly of polypeptides of the light-harvesting 2 complex encoded by distinct operons during acclimation of Rhodobacter sphaeroides to low light intensity. Photosynth Res 111, 125–138 (2012). https://doi.org/10.1007/s11120-011-9707-4

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