Characterization of csmB genes, encoding a 7.5-kDa protein of the chlorosome envelope, from the green sulfur bacteria Chlorobium vibrioforme 8327D and Chlorobium tepidum.
The csmB gene, encoding the 7.5-kDa "Gerola-Olson" protein of chlorosomes, has been cloned and sequenced from the green sulfur bacteria Chlorobium vibrioforme strain 8327D and Chlorobium tepidum. Two potential start codons were identified, and the csmB gene may be translated into a preprotein with an amino-terminal extension. Two forms of the mature CsmB protein (74 or 75 amino acids in length) were identified that differ by the presence or absence of a methionine residue at the amino terminus. The csmB gene of Chl. tepidum is transcribed as an abundant monocistronic mRNA of approximately 350 nucleotides; primer extension mapping of the 5' endpoint of the csmB mRNA suggests there is strong similarity between the csmB promoter and the sigma70 promoters of Escherichia coli. The CsmB protein of Chl. tepidum was overproduced as a histidine-tagged fusion protein in E. coli, purified to homogeneity by Ni2+ chelation affinity chromatography, and used to raise polyclonal antibodies in rabbits. Protease susceptibility mapping and agglutination experiments with isolated chlorosomes using anti-CsmB antibodies indicate that the CsmB protein is a component of the chlorosome envelope.
Chung
S
S
Department of Biochemistry and Molecular Biology, and Center for Biological Structure and Function, The Pennsylvania State University, University Park, PA 16802, USA. DAB14@PSUVM.PSU.EDU
Bryant
D A
DA
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D019414:Chlorobi / Q000235:genetics*; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D012091:Repetitive Sequences, Nucleic Acid; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; C404704:CsmB protein, Chlorobium sp.; D017931:DNA Primers; D045322:Photosynthetic Reaction Center Complex Proteins; D010450:Endopeptidases