title: The Emb proteins of mycobacteria direct arabinosylation of lipoarabinomannan and arabinogalactan via an N-terminal recognition region and a C-terminal synthetic region.
abstract: The arabinans of the mycobacterial cell wall are key structural and immunological polymers in the context of arabinogalactan (AG) and lipoarabinomannan (LAM) respectively. The three homologous membrane proteins EmbA, EmbB and EmbC are known to be involved in the synthesis of arabinan but their biochemical functions are not understood. Herein we show, that synthesis of LAM, but not AG, ceases after inactivation of embC in Mycobacterium smegmatis by insertional mutagenesis. LAM synthesis is restored upon complementation with the embC wild-type gene. Previously we have shown that the synthesis of the arabinan of AG is affected by embA or embB disruption. Thus the Emb proteins are capable of differential recognition of the galactan or mannan acceptors prior to appropriate arabinosylation. In addition, a combination of genetic and biochemical approaches have allowed us to assign some specific functions to the regions of emb gene products. Complementation of the embCmacr; mutant with a hybrid gene encoding the N-terminus of EmbC and the C-terminus of EmbB resulted in LAM with a lower molecular weight than the wild-type LAM. Structural studies involving enzyme digestion, chromatography and mass spectrometry analyses revealed that the arabinan of the 'LAM' formed in the hybrid was of AG kind rather than LAM type of arabinan.
mesh_terms: D001089:Arabinose / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D005685:Galactans / Q000096:biosynthesis* / Q000737:chemistry; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D008070:Lipopolysaccharides / Q000096:biosynthesis* / Q000737:chemistry; D016254:Mutagenesis, Insertional; D020102:Mycobacterium smegmatis / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D005685:Galactans; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; C050016:lipoarabinomannan; D001089:Arabinose; C005653:arabinogalactan