International Institute of Molecular and Cell Biology Warsaw, Poland Max-Planck Institute of...
-
Upload
angelina-wiggins -
Category
Documents
-
view
220 -
download
0
Transcript of International Institute of Molecular and Cell Biology Warsaw, Poland Max-Planck Institute of...
International Institute ofMolecular and Cell BiologyWarsaw, Poland
Max-Planck Institute ofMolecular Cell Biology and Genetics
Dresden, Germany
Laboratory of Structural BiologyMax-Planck-PAN Joint Junior Research Group
Physics in Biology:
Protein Crystallography
From physics to biology
Crystallography of sapphire
Walther group, Munich
Protein crystallography
Warsaw
10 108 10-8
Crystallography shows tremendous complexity at high magnification.
Bacterial cell walls
Gram negative Gram positive
membranemembrane
peptidoglycanpeptidoglycan
membrane
Peptidoglycan detail
N-domain catalytic domain C-domainlysostaphin
LytM
HxH
HxH
LytM was studied as a model system for lysostaphin.
LytM is a new fold in proteolysis
N-domain
C-domain
Zinc ligands:
H 210D 214H 293
Occluding ligand:
N117
S. G. Odintsov, I. Sabala, M. Marcyjaniak, and M. Bochtler, Latent LytM at 1.3A resolution. J Mol Biol, 2004. 335(3): p. 775-85.
LytM, mature form
D-Ala-D-Ala carboxypeptidase
Sonic hedgehog, N-domain
Bochtler M, Odintsov SG, Marcyjaniak M, Sabala I. Similar active sites in lysostaphins and D-Ala-D-Ala metallopeptidases. Protein Sci. 2004 13 (4): p. 854-61.
Unexpected local similarities
HX3-6D
HXH
MepA-type enzymes could be LAS enzymes.
LytM (active form) MepA
D-Ala-D-Ala
carboxypeptidase
VanX
Sonic hedgehog
N-domain
Summary: the LAS group so far
Another unknown peptidoglycan amidase, unknown fold and function
blue highest conservation, red lowest conservation
Korza HJ, Bochtler M. P. aeruginosa LD-carboxypeptidase: A serine peptidase with a Ser-His-Glu triad and a nucleophilic elbow. J Biol Chem. 2005 Sep 14; [Epub ahead of print]
A serine peptidase with a Ser-His-Glu triad.
Note that mutated enzymes were used in 100-fold higher concentration and incubated longer than wild-type enzyme to make the comparison more stringent.
The active site serines are Ramachandran outliers.
LD-carboxypeptidase
-hydrolase
(in this case, a lipase)
Acknowledgement: