Crystal structure of a novel germination protease from spores of Bacillus megaterium: structural arrangement and zymogen activation.
The DNA in the core of spores of Bacillus species is saturated with a group of small, acid-soluble proteins (SASP) that protect DNA from a variety of harsh treatments and play a major role in spore resistance and long-term spore survival. During spore germination, SASPs are rapidly degraded to amino acids and this degradation is initiated by a sequence-specific protease called germination protease (GPR), which exhibits no obvious mechanistic or amino acid sequence similarity to any known class of proteases. GPR is synthesized during sporulation as an inactive tetrameric zymogen termed P(46), which later autoprocesses to a smaller form termed P(41), which is active only during spore germination. Here, we report the crystal structure of P(46) from Bacillus megaterium at 3.0 A resolution and the fact that P(46) monomer adopts a novel fold. The asymmetric unit contains two P(46) monomers and the functional tetramer is a dimer of dimers, with an approximately 9 A channel in the center of the tetramer. Analysis of the P(46) structure and site-directed mutagenesis studies have provided some insight into the mechanism of zymogen activation as well as the zymogen's lack of activity and the inactivity of P(41) in the mature spore.
Ponnuraj
K
K
Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL, 35294, USA.
Rowland
S
S
Nessi
C
C
Setlow
P
P
Jedrzejas
M J
MJ
D000595:Amino Acid Sequence; D001410:Bacillus megaterium / Q000201:enzymology*; D018360:Crystallography, X-Ray; D019281:Dimerization; D010450:Endopeptidases / Q000737:chemistry* / Q000378:metabolism*; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D016415:Sequence Alignment; D013171:Spores, Bacterial / Q000201:enzymology*
D004792:Enzyme Precursors; D010450:Endopeptidases; C104642:spore germination protease