A Ca²⁺-binding motif underlies the unusual properties of certain photosynthetic bacterial core light-harvesting complexes

The mildly thermophilic purple phototrophic bacterium Allochromatium tepidum provides a unique model for investigating various intermediate phenotypes observed between those of thermophilic and mesophilic counterparts. The core light-harvesting (LH1) complex from A. tepidum exhibits an absorption maximum at 890 nm and mildly enhanced thermostability, both of which are Ca²⁺-dependent. However, it is unknown what structural determinants might contribute to these properties. Here, we present a cryo-EM structure of the reaction center-associated LH1 complex at 2.81 Å resolution, in which we identify multiple pigment-binding α- and β-polypeptides within an LH1 ring. Of the 16 α-polypeptides, we show that six (α1) bind Ca²⁺ along with β1- or β3-polypeptides to form the Ca²⁺-binding sites. This structure differs from that of fully Ca²⁺-bound LH1 from Thermochromatium tepidum, enabling determination of the minimum structural requirements for Ca²⁺-binding. We also identified three amino acids (Trp44, Asp47, and Ile49) in the C-terminal region of the A. tepidum α1-polypeptide that ligate each Ca ion, forming a Ca²⁺-binding WxxDxI motif that is conserved in all Ca²⁺-bound LH1 α-polypeptides from other species with reported structures. The partial Ca²⁺-bound structure further explains the unusual phenotypic properties observed for this bacterium in terms of its Ca²⁺-requirements for thermostability, spectroscopy, and phototrophic growth, and supports the hypothesis that A. tepidum may represent a "transitional" species between mesophilic and thermophilic purple sulfur bacteria. The characteristic arrangement of multiple αβ-polypeptides also suggests a mechanism of molecular recognition in the expression and/or assembly of the LH1 complex that could be regulated through interactions with reaction center subunits.