SASDA58: UL26N of pseudorabies virus (VP24, UL26N)

Basic information

• Entry: Database: SASBDB / ID: SASDA58

Sample

UL26N of pseudorabies virus

• VP24 (protein), UL26N, Suid herpesvirus 1

Function / homology

Function:

assemblin / viral release from host cell / host cell cytoplasm / serine-type endopeptidase activity / host cell nucleus / proteolysis / identical protein binding

Domain/homology:

Peptidase S21 / Herpesvirus protease superfamily / Assemblin (Peptidase family S21)

Component:

Capsid scaffolding protein

• Biological species: Suid herpesvirus 1
• Citation: Journal: PLoS Pathog / Year: 2015; Title: Dimerization-Induced Allosteric Changes of the Oxyanion-Hole Loop Activate the Pseudorabies Virus Assemblin pUL26N, a Herpesvirus Serine Protease.; Authors: Martin Zühlsdorf / Sebastiaan Werten / Barbara G Klupp / Gottfried J Palm / Thomas C Mettenleiter / Winfried Hinrichs / ; Abstract: Herpesviruses encode a characteristic serine protease with a unique fold and an active site that comprises the unusual triad Ser-His-His. The protease is essential for viral replication and as such constitutes a promising drug target. In solution, a dynamic equilibrium exists between an inactive monomeric and an active dimeric form of the enzyme, which is believed to play a key regulatory role in the orchestration of proteolysis and capsid assembly. Currently available crystal structures of herpesvirus proteases correspond either to the dimeric state or to complexes with peptide mimetics that alter the dimerization interface. In contrast, the structure of the native monomeric state has remained elusive. Here, we present the three-dimensional structures of native monomeric, active dimeric, and diisopropyl fluorophosphate-inhibited dimeric protease derived from pseudorabies virus, an alphaherpesvirus of swine. These structures, solved by X-ray crystallography to respective resolutions of 2.05, 2.10 and 2.03 Å, allow a direct comparison of the main conformational states of the protease. In the dimeric form, a functional oxyanion hole is formed by a loop of 10 amino-acid residues encompassing two consecutive arginine residues (Arg136 and Arg137); both are strictly conserved throughout the herpesviruses. In the monomeric form, the top of the loop is shifted by approximately 11 Å, resulting in a complete disruption of the oxyanion hole and loss of activity. The dimerization-induced allosteric changes described here form the physical basis for the concentration-dependent activation of the protease, which is essential for proper virus replication. Small-angle X-ray scattering experiments confirmed a concentration-dependent equilibrium of monomeric and dimeric protease in solution.

Contact author

• Sebastiaan Werten (Ernst-Moritz-Arndt-Universität Greifswald, Domstraße 11 17489 Greifswald)

Models

• Model #311: ; Type: atomic / Software: Crysol / Chi-square value: 1.288; Search similar-shape structures of this assembly by Omokage search (details)
• Model #350: ; Type: dummy / Software: DAMMIN / Radius of dummy atoms: 2.25 A / Chi-square value: 0.902; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: UL26N of pseudorabies virus / Specimen concentration: 9.83 mg/ml
• Buffer: Name: Tris/HCl / Concentration: 50.00 mM / pH: 7.5 ; Composition: 0.5 M NaCl, 0.25 M imidazole, 5% glycerol, 50 mM urea, 0.2 M MgCl2

Entity #177

Name: UL26N / Type: protein / Description: VP24 / Formula weight: 26.608 / Num. of mol.: 2 / Source: Suid herpesvirus 1 / References: UniProt: Q83417
Sequence:

MGSSHHHHHH SSGLVPRGSH MGPVYVSGYL ALYDRDGGEL ALTREIVAAA LPPAGPLPIN IDHRPRCDIG AVLAVVDDDR GPFFLGVVNC PQLGAVLARA VGPDFFGDMR LSDEERLLYL LSNYLPSASL SSRRLAPGEA PDETLFAHVA LCVIGRRVGT IVVYDASPEA AVAPFRQLSA RARSELLARA AESPDRERVW HMSEEALTRA LLSTAVNNML LRDRWELVAA RRREAGVRGH TYLQ

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: UL26N of pseudorabies virus / Measurement date: Sep 23, 2013 / Cell temperature: 10 °C / Exposure time: 0.05 sec. / Number of frames: 20 / Unit: 1/nm /

	Min	Max
Q	0.068	4.4372

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 905 /

	Min	Max
Q	0.096949	2.47635
P(R) point	1	905
R	0	10.56

Result

D max: 10.6 / Type of curve: single_conc /

	Experimental	Porod
MW	45 kDa	45 kDa
Volume	-	73 nm3

	P(R)	Guinier
Forward scattering, I0	2982	2964
Radius of gyration, Rg	2.69 nm	2.62 nm