| Title: | Isolation of constitutive variants of a subfamily 10 histidine protein kinase (SppK) from Lactobacillus using random mutagenesis |
| Author(s): | Mathiesen G; Axelsen GW; Axelsson L; Eijsink VG; |
| Address: | "Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Chr. M. Falsensvei 1, PO Box 5003, 1432 As, Norway" |
| DOI: | 10.1007/s00203-005-0049-5 |
| ISSN/ISBN: | 0302-8933 (Print) 0302-8933 (Linking) |
| Abstract: | "The histidine protein kinase SppK is a peptide pheromone-activated kinase that regulates the production of the bacteriocin sakacin P in Lactobacillus sakei. SppK belongs to subfamily 10 of histidine protein kinases (HPKs), which regulate important processes in Gram-positive bacteria, including virulence, competence and bacteriocin production. To obtain insight into the functional properties of this relatively unknown class of HPKs, we have subjected SppK to random mutagenesis by error-prone PCR, followed by selection for mutants displaying a constitutive phenotype. Most identified mutations were clustered in a predicted coiled coil-like region, which is an important part of the HPK dimer interface and which includes the autophosphorylated histidine. Other mutations were located in the junctions between the dimerization domain and the membrane receptor domain or the catalytic kinase domain. Interestingly, two previously identified constitutive variants of ComD, an SppK homologue involved in competence regulation in Streptococcus pneumoniae, contained single mutations in the same regions" |
| Keywords: | "Amino Acid Sequence Bacteriocins/biosynthesis Dimerization Gene Expression Regulation, Bacterial *Genetic Variation Histidine Kinase Lactobacillus/*enzymology/genetics Molecular Sequence Data *Mutagenesis Mutation Polymerase Chain Reaction/methods Protein;" |
| Notes: | "MedlineMathiesen, Geir Axelsen, Gunnhild W Axelsson, Lars Eijsink, Vincent G H eng Research Support, Non-U.S. Gov't Germany 2005/11/12 Arch Microbiol. 2006 Jan; 184(5):327-34. doi: 10.1007/s00203-005-0049-5. Epub 2005 Nov 10" |
