Prediction of avian Influenza A binding preference to human receptor using

conformational analysis of receptor bound to hemagglutinin

Wanwimon Mokmak, M.Sc.Biostatistics and Informatics Laboratory

Genome Institute, BIOTEC, Thailand

TYR191TYR191

SER129SER129

GLN222GLN222 TRP149TRP149TYR91TYR91

SER133SER133

Ha, Y., et al. Proc.Natl.Acad.Sci.USA 98: 11181-11186

Chain A of H5 HA (A/Duck/Singapore/3/97 HA)

Avian influenza H5N1

Hemagglutinin protein

Host cell receptor

Introduction

Hemagglutinin Protein

potentially changing the receptor binding preference

HA Mutation

Φ = 55 degreeCis conformation

Φ = -55 degreeTran conformation

Introduction

Viseshakul N et al.Virology 2004, 328, 169-76

Avian Influenza A virus

Mutated KAN-1Mutated Sing-97

Avian receptor Human receptor

• To predict the HA binding selectivity from different HA variants in receptor-based conformational analysis by measuring a single torsion angle (Φ) during molecular dynamic simulation

Objectives

Material & Method

X-ray crystal structures 1JSO template for Sing-97 and Kan1 variant

1RVZ template for Puerto 34 variant

host cell receptor X-ray structure or

3D- homology models of HA protein

HA complex

MD simulation Binding preference

Observed torsion angleof host cell receptor

Known HA variants * :•Sing-97 •Puerto-34 (human flu virus)•Sing-97 variant (Q222L, G224S, Q222L/G224S)•Kan-1•Double KAN-1 mutant (L129V/A134V)

Prediction for HA mutants • Single Kan-1 HA : M226T, K189G, K218S and L109P• Puerto-34 HA variants :Q222L, G224L, Q222L/G224S

Comparison of observed torsion angle from binding simulation

Results & Discussion

Puerto-34

Sing-97

Kan-1

tran

cis

tran

cis

tran

cis

• Sing-97 bound better toAvian > human• Q222L/G224L sing-97bind preferentiallywith human type

• Kan-1bind better toAvian > human• L129V/A134V Kan-1bind preferentiallywith human type

• Puerto-34 bind preferentiallywith human type

Sing-97Q222L Sing-97G224S Sing-97Q222L/G224S Sing-97

Kan-1L129V/A134V KAN-1

*Harvey R et al. J Virol 2004, 78:502-507. Auewarakul P. et al. J Virol 2007, 81:9950-9955.

*

Human receptor

Avian receptor

*

Results & Discussion

MD predicted receptor type selectivity of HAs compared with data from published experimental assays

Order of binding selectivity to human receptor for flu virusesPuerto-34 > L129V/A134V Kan-1≅ Q222L/G224S Sing-97 > Sing-97 ≅ Kan-1

Tested HA variants Simulation in vitro

Puerto-34 +++ +++

Sing-97 ++ +

Q222L Sing-97 ++ ++

G224S Sing-97 + ++

Q222L/G224S Sing-97 +++ +++

Kan-1 + +

L129V/A134V Kan-1 +++ +++

+++ most preferred, ++ moderately preferred, + least preferred

Simulation results of four Kan-1 HA mutant variants binding to Human receptor

Prediction of the Relative Binding Selectivity

Results & Discussion

Tran

Cis

Simulation results of four Puerto-34 HA mutants

Prediction of the Relative Binding Selectivity

Results & Discussion

Tran

Cis

Human receptor associates with positive +Φ angle (Cis conformation)

Avian-type receptor associates with negative -Φ angle (Tran conformation)

Using Molecular dynamics simulation to observe torsion angles

Conclusions

Dr. Sissades TongsimaDr. Chak SangmaMs.Nipa JongkonMs.Daungmanee ChuakheawMr. Anunchai AssawamakinDr. Philip J. ShawDr. Jittima Piriyapongsa

Acknowledgements

National Center for Genetic Engineering and Biotechnology(BIOTEC)Kasetsart University

THANK YOU