SASDAE7: Structure of a complex between full length and truncated CS74L en...

Basic information

• Entry: Database: SASBDB / ID: SASDAE7

Sample

Structure of a complex between full length and truncated CS74L endolysin

• Endolysin CS74L (protein), CS74L, Clostridium phage phi8074-B1

• Function / homology: Ami_3 / N-acetylmuramoyl-L-alanine amidase, catalytic domain / N-acetylmuramoyl-L-alanine amidase / N-acetylmuramoyl-L-alanine amidase activity / viral release from host cell by cytolysis / peptidoglycan catabolic process / Endolysin CS74L
• Biological species: Clostridium phage phi8074-B1 (virus)
• Citation: Journal: J Biol Chem / Year: 2016; Title: Crystal Structure of the CTP1L Endolysin Reveals How Its Activity Is Regulated by a Secondary Translation Product.; Authors: Matthew Dunne / Stefan Leicht / Boris Krichel / Haydyn D T Mertens / Andrew Thompson / Jeroen Krijgsveld / Dmitri I Svergun / Natalia Gómez-Torres / Sonia Garde / Charlotte Uetrecht / Arjan Narbad / Melinda J Mayer / Rob Meijers / ; Abstract: Bacteriophages produce endolysins, which lyse the bacterial host cell to release newly produced virions. The timing of lysis is regulated and is thought to involve the activation of a molecular switch. We present a crystal structure of the activated endolysin CTP1L that targets Clostridium tyrobutyricum, consisting of a complex between the full-length protein and an N-terminally truncated C-terminal cell wall binding domain (CBD). The truncated CBD is produced through an internal translation start site within the endolysin gene. Mutants affecting the internal translation site change the oligomeric state of the endolysin and reduce lytic activity. The activity can be modulated by reconstitution of the full-length endolysin-CBD complex with free CBD. The same oligomerization mechanism applies to the CD27L endolysin that targets Clostridium difficile and the CS74L endolysin that targets Clostridium sporogenes. When the CTP1L endolysin gene is introduced into the commensal bacterium Lactococcus lactis, the truncated CBD is also produced, showing that the alternative start codon can be used in other bacterial species. The identification of a translational switch affecting oligomerization presented here has implications for the design of effective endolysins for the treatment of bacterial infections.

Contact author

• Haydyn Mertens (EMBL-Hamburg, European Molecular Biology Laboratory (EMBL) - Hamburg outstation, Notkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany)

Models

• Model #257: ; Type: dummy / Radius of dummy atoms: 3.00 A / Chi-square value: 0.775; Search similar-shape structures of this assembly by Omokage search (details)
• Model #258: ; Type: mix / Software: (2.0) / Radius of dummy atoms: 1.90 A / Comment: Ensemble Component: Heterotetramer (compact) / Chi-square value: 0.664 / P-value: 0.051000; Search similar-shape structures of this assembly by Omokage search (details)
• Model #259: ; Type: mix / Radius of dummy atoms: 1.90 A / Comment: Ensemble Component: Heterotetramer (extended) / Chi-square value: 0.664 / P-value: 0.051000; Search similar-shape structures of this assembly by Omokage search (details)
• Model #260: ; Type: mix / Software: (2.0) / Radius of dummy atoms: 1.90 A / Comment: Ensemble Component: Free CTD of endolysin / Chi-square value: 0.664 / P-value: 0.051000; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: Structure of a complex between full length and truncated CS74L endolysin; Specimen concentration: 1.00-6.80
• Buffer: Name: 20 mM HEPES / pH: 7.4

Entity #156

Name: CS74L / Type: protein / Description: Endolysin CS74L / Formula weight: 31.1 / Source: Clostridium phage phi8074-B1 / References: UniProt: I1TJX3
Sequence:

MGSSHHHHHH SSGLVPRGSH MKIGIDMGHT LSGADYGVVG LRPESVLTRE VGTKVIYKLQ KLGHVVVNCT VDKASSVSES LYTRYYRANQ ANVDLFISIH FNATPGGTGT EVYTYAGRQL GEATRIRQEF KSLGLRDRGT KDGSGLAVIR NTKAKAMLVE CCFCDNPNDM KLYNSESFSN AIVKGITGKL PNGESGNNNQ GGNKVKAVVI YNEGADRRGA EYLADYLNCP TISNSRTFDY SCVEHVYAVG GKKEQYTKYL KTLLSGANRY DTMQQILNFI NGGK

Experimental information

• Beam: Instrument name: DORIS III X33 / City: Hamburg / 国: Germany / Shape: 0.6 / Type of source: X-ray synchrotron / Wavelength: 0.15 Å / Dist. spec. to detc.: 2.7 mm
• Detector: Name: Pilatus 1M-W / Pixsize x: 0.172 mm

Scan

Title: CS74L Endolysin / Measurement date: Mar 17, 2011 / Cell temperature: 20 °C / Exposure time: 15 sec. / Number of frames: 8 / Unit: 1/nm /

	Min	Max
Q	0.1628	6.0241

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 315 /

	Min	Max
Q	0.2845	2.004
P(R) point	1	315
R	0	14

Result

D max: 140 / Type of curve: merged

	Experimental	Standard	Porod
MW	38.4 kDa	38.4 kDa	49.5 kDa
Volume	-	-	79 nm3

	P(R)	Guinier
Forward scattering, I0	51.9	50.3
Radius of gyration, Rg	3.8 nm	3.6 nm