SASDB74: CyaA Block I-V (Adenylate cyclase toxin Block I-V)

Basic information

• Entry: Database: SASBDB / ID: SASDB74

Sample

CyaA Block I-V

• Adenylate cyclase toxin Block I-V (protein), CyaA Block I-V, Bordetella pertussis

Function / homology

Function:

calcium- and calmodulin-responsive adenylate cyclase activity / hemolysis in another organism / adenylate cyclase / cAMP biosynthetic process / adenylate cyclase activity / channel activity / positive regulation of cytosolic calcium ion concentration / toxin activity / calmodulin binding / calcium ion binding / host cell plasma membrane / extracellular region / ATP binding / membrane

Domain/homology:

RTX, pore-forming domain / N-terminal domain in RTX protein / Haemolysin-type calcium binding-related / Haemolysin-type calcium binding protein related domain / RTX toxin determinant A / Anthrax toxin, edema factor, central / Anthrax toxin, edema factor, C-terminal / Anthrax toxin, edema factor, central domain superfamily / Anthrax toxin LF subunit / Hemolysin-type calcium-binding conserved site / Hemolysin-type calcium-binding region signature. / RTX calcium-binding nonapeptide repeat / RTX calcium-binding nonapeptide repeat (4 copies) / Serralysin-like metalloprotease, C-terminal

Component:

Bifunctional hemolysin/adenylate cyclase

• Biological species: Bordetella pertussis (bacteria)
• Citation: Journal: Mol Cell / Year: 2016; Title: Calcium-Driven Folding of RTX Domain β-Rolls Ratchets Translocation of RTX Proteins through Type I Secretion Ducts.; Authors: Ladislav Bumba / Jiri Masin / Pavel Macek / Tomas Wald / Lucia Motlova / Ilona Bibova / Nela Klimova / Lucie Bednarova / Vaclav Veverka / Michael Kachala / Dmitri I Svergun / Cyril Barinka / Peter Sebo / ; Abstract: Calcium-binding RTX proteins are equipped with C-terminal secretion signals and translocate from the Ca(2+)-depleted cytosol of Gram-negative bacteria directly into the Ca(2+)-rich external milieu, passing through the "channel-tunnel" ducts of type I secretion systems (T1SSs). Using Bordetella pertussis adenylate cyclase toxin, we solved the structure of an essential C-terminal assembly that caps the RTX domains of RTX family leukotoxins. This is shown to scaffold directional Ca(2+)-dependent folding of the carboxy-proximal RTX repeat blocks into β-rolls. The resulting intramolecular Brownian ratchets then prevent backsliding of translocating RTX proteins in the T1SS conduits and thereby accelerate excretion of very large RTX leukotoxins from bacterial cells by a vectorial "push-ratchet" mechanism. Successive Ca(2+)-dependent and cosecretional acquisition of a functional RTX toxin structure in the course of T1SS-mediated translocation, through RTX domain folding from the C-terminal cap toward the N terminus, sets a paradigm that opens for design of virulence inhibitors of major pathogens.

Contact author

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

Models

• Model #484: ; Type: dummy / Software: DAMMIF / Radius of dummy atoms: 4.75 A / Chi-square value: 0.839056; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: CyaA Block I-V / Specimen concentration: 1.10-4.40
• Buffer: Name: Tris HCl / Concentration: 10.00 mM / pH: 8 / Composition: 150 mM NaCl, 10 mM CaCl2

Entity #316

Name: CyaA Block I-V / Type: protein / Description: Adenylate cyclase toxin Block I-V / Formula weight: 70 / Num. of mol.: 1 / Source: Bordetella pertussis / References: UniProt: P0DKX7
Sequence:

EGDIHMSPIL GYWKIKGLVQ PTRLLLEYLE EKYEEHLYER DEGDKWRNKK FELGLEFPNL PYYIDGDVKL TQSMAIIRYI ADKHNMLGGC PKERAEISML EGAVLDIRYG VSRIAYSKDF ETLKVDFLSK LPEMLKMFED RLCHKTYLNG DHVTHPDFML YDALDVVLYM DPMCLDAFPK LVCFKKRIEA IPQIDKYLKS SKYIAWPLQG WQATFGGGDH PPKSDGSTSG SGHHHHHHSA GLVPRGSTAI GMKETAAAKF ERQHMDSPDL GTGGGSGIEG Renlyfqgle hvqhiiggag ndsitgnahd nflaggsgdd rldggagndt lvggegqntv iggagddvfl qdlgvwsnql dggagvdtvk ynvhqpseer lermgdtgih adlqkgtvek wpalnlfsvd hvknienlhg srlndriagd dqdnelwghd gndtirgrgg ddilrgglgl dtlygedgnd iflqddetvs ddidggagld tvdysamihp grivapheyg fgieadlsre wvrkasalgv dyydnvrnve nvigtsmkdv ligdaqantl mgqggddtvr ggdgddllfg gdgndmlygd agndtlyggl gddtleggag ndwfgqtqar ehdvlrggdg vdtvdysqtg ahagiaagri glgiladlga grvdklgeag ssaydtvsgi envvgtelad ritgdaqanv lrgaggadvl aggegddvll ggdgddqlsg dagrdrlyge agddwffqda anagnlldgg dgrdtvdfsg pgrgldagak gvflslgkgf aslmdepets nvlrnienav gsarddvlig daganvlngl agndvlsgga gddvllgdeg sdllsgdagn ddlfggqgdd tylfgvgygh dtiyesgggh dtirinagad qlwfarqgnd leirilgtdd altvhdwyrd adhrveiiha anqavdqagi eklveamaqy pdp

Experimental information

• Beam: Instrument name: PETRA III P12 / City: Hamburg / 国: Germany / Type of source: X-ray synchrotronSynchrotron / Wavelength: 0.12 Å / Dist. spec. to detc.: 3.1 mm
• Detector: Name: Pilatus 2M

Scan

Title: CyaA Block I-V / Measurement date: Oct 31, 2013 / Storage temperature: 10 °C / Cell temperature: 10 °C / Exposure time: 0.05 sec. / Number of frames: 20 / Unit: 1/nm /

	Min	Max
Q	0.0291	4.4884

Distance distribution function P(R)

Sofotware P(R): GNOM 4.6 / Number of points: 503 /

	Min	Max
Q	0.1002	1.422
P(R) point	28	530
R	0	17.15

Result

D max: 17.15 / Type of curve: extrapolated / Standard: BSA

	Experimental	Standard	Porod
MW	91 kDa	91 kDa	162 kDa
Volume	-	-	275 nm3

	P(R)	Guinier
Forward scattering, I0	17070	17551
Radius of gyration, Rg	5.49 nm	5.55 nm