SASDC63: Fowlpox Virus FPV039 antiapoptotic Bcl-2 viral protein in complex...

Basic information

Entry

Database: SASBDB / ID: SASDC63

Sample

Fowlpox Virus FPV039 antiapoptotic Bcl-2 viral protein in complex with BaK BH3 peptide (FPV039:BAK)

• Bcl-2-like protein FPV039 (protein), FPV039, Fowlpox virus (strain NVSL)
• Uncharacterized protein (BAK1) (protein), Bak, Gallus gallus

Function / homology

Function:

Activation and oligomerization of BAK protein / Release of apoptotic factors from the mitochondria / Pyroptosis / BH domain binding / symbiont-mediated suppression of host apoptosis / porin activity / host cell mitochondrion / extrinsic apoptotic signaling pathway in absence of ligand / release of cytochrome c from mitochondria / intrinsic apoptotic signaling pathway in response to DNA damage / regulation of apoptotic process / mitochondrial outer membrane

Domain/homology:

Apoptosis regulator, Bcl-2, BH1 motif, conserved site / Apoptosis regulator, Bcl-2 family BH1 motif signature. / Apoptosis regulator, Bcl-2, BH2 motif, conserved site / Apoptosis regulator, Bcl-2 family BH2 motif signature. / Bcl-2 family / BCL (B-Cell lymphoma); contains BH1, BH2 regions / Bcl2-like / Bcl-2, Bcl-2 homology region 1-3 / Apoptosis regulator proteins, Bcl-2 family / BCL2-like apoptosis inhibitors family profile. / Bcl-2-like superfamily

Component:

Bcl-2 Bcl-2 homology region 1-3 domain-containing protein / Apoptosis regulator Bcl-2 homolog

• Biological species: Fowlpox virus (strain NVSL); Gallus gallus (chicken)
• Citation: Journal: J Biol Chem / Year: 2017; Title: Structural basis of apoptosis inhibition by the fowlpox virus protein FPV039.; Authors: Mohd Ishtiaq Anasir / Sofia Caria / Michael A Skinner / Marc Kvansakul / ; Abstract: Programmed cell death or apoptosis of infected host cells is an important defense mechanism in response to viral infections. This process is regulated by proapoptotic and prosurvival members of the B-cell lymphoma 2 (Bcl-2) protein family. To counter premature death of a virus-infected cell, poxviruses use a range of different molecular strategies including the mimicry of prosurvival Bcl-2 proteins. One such viral prosurvival protein is the fowlpox virus protein FPV039, which is a potent apoptosis inhibitor, but the precise molecular mechanism by which FPV039 inhibits apoptosis is unknown. To understand how fowlpox virus inhibits apoptosis, we examined FPV039 using isothermal titration calorimetry, small-angle X-ray scattering, and X-ray crystallography. Here, we report that the fowlpox virus prosurvival protein FPV039 promiscuously binds to cellular proapoptotic Bcl-2 and engages all major proapoptotic Bcl-2 proteins. Unlike other identified viral Bcl-2 proteins to date, FPV039 engaged with cellular proapoptotic Bcl-2 with affinities comparable with those of Bcl-2's endogenous cellular counterparts. Structural studies revealed that FPV039 adopts the conserved Bcl-2 fold observed in cellular prosurvival Bcl-2 proteins and closely mimics the structure of the prosurvival Bcl-2 family protein Mcl-1. Our findings suggest that FPV039 is a pan-Bcl-2 protein inhibitor that can engage all host BH3-only proteins, as well as Bcl-2-associated X, apoptosis regulator (Bax) and Bcl-2 antagonist/killer (Bak) proteins to inhibit premature apoptosis of an infected host cell. This work therefore provides a mechanistic platform to better understand FPV039-mediated apoptosis inhibition.

Contact author

• Sofia Caria

Models

Sample

• Sample: Name: Fowlpox Virus FPV039 antiapoptotic Bcl-2 viral protein in complex with BaK BH3 peptide (FPV039:BAK); Specimen concentration: 0.31-7.35 / Entity id: 631 / 632
• Buffer: Name: 20 mM trisodium citrate pH, 200 mM NaCl / pH: 6

Entity #631

Name: FPV039 / Type: protein / Description: Bcl-2-like protein FPV039 / Formula weight: 16.823 / Num. of mol.: 1 / Source: Fowlpox virus (strain NVSL) / References: UniProt: Q9J5G4
Sequence:

MASSNMKDET YYIALNMIQN YIIEYNTNKP RKSFVIDSIS YDVLKAACKS VIKTNYNEFD IIISRNIDFN VIVTQVLEDK INWGRIITII AFCAYYSKKV KQDTSPQYYD GIISEAITDA ILSKYRSWFI DQDYWNGIRI YKN

Entity #632

Name: Bak / Type: protein / Description: Uncharacterized protein (BAK1) / Formula weight: 2.805 / Num. of mol.: 1 / Source: Gallus gallus / References: UniProt: Q5F404
Sequence:

LGSTGSQVGR RLAIIGDDIN KRYDAE

Experimental information

• Beam: Instrument name: Australian Synchrotron SAXS/WAXS / City: Melbourne / 国: Australia / Shape: Point / Type of source: X-ray synchrotron / Dist. spec. to detc.: 1.6 mm
• Detector: Name: Pilatus 1M

Scan

Title: Fowlpox Virus FPV039 antiapoptotic Bcl-2 viral protein in complex with BaK BH3 peptide (FPV039:BAK)
Measurement date: Oct 2, 2015 / Storage temperature: 20 °C / Cell temperature: 20 °C / Exposure time: 1 sec. / Number of frames: 30 / Unit: 1/A /

	Min	Max
Q	0.0106	0.6185

Result

Experimental MW: 17.9 kDa / Type of curve: single_conc

	Guinier	Guinier error
Forward scattering, I0	0.104031	0.0002
Radius of gyration, Rg	1.96 nm	0.006

	Min	Max
Guinier point	38	70