Proteome analysis of the model microsymbiont Sinorhizobium meliloti: Isolation and characterisation of novel proteins

Abstract

Sinorhizobium meliloti is an agriculturally and ecologically important microbe due to its capacity to establish nitrogen-fixing symbiosis with plant legumes. Two-dimensional gel electrophoresis of total cellular protein was used to establish a proteome reference map for the model microsymbiont Sinorhizobium meliloti strain 1021. The extent of changes in the gene expression of cells grown in a defined medium at different growth phases was established. After examination of over 2000 resolved protein spots, a minimum of 52 reproducible changes in protein expression levels were detected when early exponential phase cells were compared to late exponential phase cells. In contrast, induction of nodulation gene expression by the addition of the flavonoid luteolin to cells did not result in detectable changes in protein expression at either early or late exponential phase. N-terminal microsequencing of eighteen unknown constitutive proteins plus four proteins, induced or up-regulated in late exponential phase cells, allowed the identification of proteins not previously described in rhizobia. These included an amide-binding protein, a putative hydrolase of the glyoxalase II protein family, a nucleoside diphosphate kinase, and a 5'-nucleotidase. N- terminal microsequencing was also valuable in revealing N-terminal post- translational processing and assigning a subcellular location to the analysed protein. Proteome analysis will provide a powerful analytical tool to complement the sequencing of the genome of strain 1021.