Shuishu Wang, PhD

Shuishu Wang, PhD

shuishu wang

Name: Shuishu Wang, PhD

Department of Primary Appointment: Biochemistry
Position: USU Faculty
Title: Assistant Professor

Email: shuishu.wang@usuhs.edu (link sends e-mail)
Office Phone: (301) 295-3418
Lab Phone: (301) 295-3111
Room: B4012

Links
Department Website

Profile

Profile

  • Ph.D. Purdue University
  • Postdoc University of California, Los Angeles

Research interests

1. PhoP-PhoR two-component signaling system of Mycobacterium tuberculosis

The PhoP-PhoR two-component system is essential for virulence of M. tuberculosis. M. tuberculosis mutants with either phoP or phoR gene disrupted have defects in cell envelope and cannot grow in human and mouse macrophages or in mice. PhoR is a sensor histidine kinase that senses environmental signals. The signals are passed onto PhoP, a response regulator that functions as a transcription regulator, through phosphorylation of its N-terminal regulatory domain to regulate its activities. We are using X-ray crystallography to study the structure and function of this two-component system.

The crystal structure of PhoP reveals two distinct structural domains, an N-terminal receiver domain that contains the phosphorylation site aspartate and a C-terminal DNA-binding domain. The two domains are tethered together through a flexible linker. The sequence recognition helix is exposed, consistent with the observation that PhoP with or without phosphorylation is able to bind DNA. Phosphorylated PhoP has a higher binding affinity to DNA. PhoP binds to direct-repeat DNA sequences of 7 bp motifs with a spacer of 4 bp as a dimer. The crystal structure of a PhoP-DNA complex reveals the details of the interactions between PhoP and DNA and between the two PhoP subunits. The two subunits binds DNA in tandem, with an asymmetric subunit-subunit interface that involves both receiver and effector domains. Because PhoP binds DNA essentially only as a dimer, the dimer interface is an ideal site for inhibitors to bind to disrupt the PhoP function. Guided by the structure of the PhoP-DNA complex, we designed a high-throughput screening platform to search for inhibitors of PhoP-DNA binding interactions.

PhoR has a modular domain structure: an extracytosolic sensor domain, a transmembrane domain, and a cytosolic domain. The cytosolic domain can be subdivided into a HAMP domain, a dimerization domain, and an ATP-binding domain. A truncated domain containing both dimerization and ATP-binding domains is functional as a kinase and can autophosphorylate and phosphorylate PhoP, although the entire cytosolic domain has a much higher activity. We have obtained crystals of most of these isolated domains, and have determined a crystal structure of the dimerization domain, which reveals an asymmetric dimeric structure. The connectivity of the helical bundle of the dimerization domain indicates that PhoR is likely to autophosphorylate in cis.

2. Structure-based design of immunogens against gonorrhea

Gonorrhea has a major impact on reproductive and neonatal health and is a common infection with an estimated 106 million new cases each year worldwide. Alarmingly, treatment options for gonorrhea are quickly diminishing with the emergence of Neisseria gonorrhoeae strains that are resistant to the last-line antibiotics. A gonorrhea vaccine will be likely the best long-term solution to this problem. We are using structure-based design to develop gonorrhea vaccines, in collaboration with Dr. Ann Jerse in the Microbiology Department of USUHS. Dr. Jerse and others have identified N. gonorrhoeae antigens, such as MtrE and PorB, which are outer-membrane β-barrel proteins. These proteins have short antigenic surface-exposed loops that are extended from the membrane-buried β-strands. Our approach is to engineer nanoparticles that display these antigenic loops on the surface in a way that maintains the native conformation of the loops. We use the Helicobacter pylori ferritin as a nanoparticle platform.

 

Selected publications

L. Wang, D. Xing, A. Le Van, A.E. Jerse, and S. Wang. Structure-based design of ferritin-nanoparticle immunogens displaying antigenic loops of Neisseria gonorrhoeae. FEBS Open Bio (2017) In press. DOI: 10.1002/2211-5463.12267

D. Xing, M.B. Ryndak, L.Wang, I. Kolesnikova, I. Smith, and S. Wang. Asymmetric structure of the dimerization domain of PhoR, a sensor kinase important for the virulence of Mycobacterium tuberculosis. ACS Omega (2017) 2, 3509-17. http://doi.org/10.1021/acsomega.7b00612

L. Wang, M. Xu, N. Southall, W. Zheng, and S. Wang. A High-Throughput Assay for Developing Inhibitors of PhoP, A Virulence Factor of Mycobacterium tuberculosis. Comb Chem High Throughput Screen. (2016) 19, 855-864.

X. He, L. Wang, and S. Wang. Structural basis of DNA sequence recognition by the response regulator PhoP in Mycobacterium tuberculosis. Sci Rep. (2016) 6, 24442. https://www.nature.com/articles/srep24442

X. He and S. Wang. DNA Consensus Sequence Motif for Binding Response Regulator PhoP, a Virulence Regulator of Mycobacterium tuberculosis. Biochemistry (2014), 53, 8008-20.

N. Sambuughin, X. Liu, S. Bijarnia, T. Wallace, I.C. Verma, S. Hamilton, S. Muldoon, L.J. Tallon and S. Wang. Exome sequencing reveals SCO2 mutations in a family presented with fatal infantile hyperthermia. J. Hum. Genet. (2013) 58, 226-8.

S. Wang. Bacterial Two-Component Systems: Structures and Signaling Mechanisms, in Protein Phosphorylation in Human Health, p439-466, (2012), C. Huang (Ed.), ISBN: 978-953-51-0737-8, InTech, DOI: 10.5772/48277. http://www.intechopen.com/articles/show/title/bacterial-two-component-sy...

S. Menon and S. Wang. Structure of the response regulator PhoP from Mycobacterium tuberculosis reveals a dimer through the receiver domain. Biochemistry (2011), 50, 5948-5957.

M. B. Ryndak, S. Wang, I. Smith, and G. M. Rodriguez. The Mycobacterium tuberculosis high-affinity iron importer, IrtA, contains an FAD-binding Domain. J. Bacteriol. (2010) 192, 861-869.

M. Ryndak, S. Wang, and I. Smith. PhoP, a key player in Mycobacterium tuberculosis virulence. Trends in Microbiology (2008) 16, 861-869.

S. Wang, J. Engohang-Ndong, and I. Smith. Structure of the DNA-binding domain of the response regulator PhoP from Mycobacterium tuberculosis. Biochemistry (2007) 46, 14751-14761.

S. Wang and D. Eisenberg. Crystal Structure of the Pantothenate Synthetase from Mycobacterium tuberculosis, Snapshots of the Enzyme in Action. Biochemistry (2006) 45, 1554-1561.

M. Strong, M. Sawaya, S. Wang, M. Phillips, D. Cascio and D. Eisenberg. Toward the structural genomics of complexes: crystal structure of a PE/PPE protein complex from Mycobacterium tuberculosis. Proc Natl Acad Sci U S A. (2006) 103, 8060-8065.

S. Wang and D. Eisenberg. Crystal Structures of a Pantothenate Synthetase from M. tuberculosis and its complexes with substrates and a reaction intermediate. Protein Science (2003) 12, 1097-1108.

S. Wang, C. Mura, M. Sawaya, D. Cascio and D. Eisenberg. Crystal Structure of a Nudix Protein from Pyrobaculum aerophilum reveals a dimer with two intersubunit β-sheets. Acta Cryst. (2002) D58, 571-578.

S. Wang, L. Tabernero, M. Zhang, E. Harms, R. L. Van Etten and C. V. Stauffacher. Crystal Structures of a Low Molecular Weight Protein Tyrosine Phosphatase from Saccharomyces cerevisiae and its Complex with the Substrate p-Nitrophenyl Phosphate. Biochemistry (2000) 39, 1903-1914.

S. Wang, C. V. Stauffacher and R. L. Van Etten. Structural and Mechanistic Basis for the Activation of a Low Molecular Weight Protein Tyrosine Phosphatase by Adenine. Biochemistry (2000) 39, 1234-1242.

 

Patent applications

D. Eisenberg, S. Wang and C. Goulding (2006) Anti-Tuberculosis Drugs Based on Pantothenate Synthetase Inhibitors. U.S. Provisional Application No. 60/757,000.

A. Jerse, M. Bash, and S. Wang (2012) Cyclic Porin Loop Peptide Gonorrhea Vaccine. U.S. provisional patent application.

A. Jerse and S. Wang (2017) Ferritin-Based Nanoparticle Vaccine Against Gonorrhea. U.S. Provisional Patent Application No. 62/526,308.