Research Interest 

We use mass spectrometric and proteomic approaches to study the molecular mechanisms of metal ion homeostasis, oxidative stress as well as their relevance to neurodegenerative diseases and cancer.

Research Interest 1. We have been developing simple and effective mass spectrometric approaches for proteomic studies. Particular interests are to identify post-translational modifications involved in signal transduction and to quantitate protein expression levels at the genomic scale.  

Research Interest 2. We have employed the proteomic approaches to carry out studies of metal homeostasis in the model organism S. cerevisiae. This will involve metal ion uptake, transport, utilization and storage as well as the relevant gene regulation required to maintain appropriate metal ion levels in vivo.  These studies are of great significance because metal ions play critical roles in cellular functions and altered metal metabolism has been related to a variety of diseases, especially neurodegenerative diseases. The focuses of the research are: analyze proteomes of yeast strains with and without specific metal ion regulation genes; and identify novel proteins involved in metal ion metabolism.  The most recent project is to study the significance of zinc homeostasis in prostate tissue and the pathogenesis of prostate cancer.

Research Interest 3. We also carry out proteomic studies of amyotrophic lateral sclerosis (ALS, Lou Gehrig’s disease). Mutations in copper-zinc superoxide dismutase (SOD1) have been linked to a subset of familial ALS, but the etiology remains largely unclear. By studying the proteomes of motor neurons expressing wild-type SOD1 and ALS-associated mutants, we expect to achieve a genome-wide understanding of protein molecules in motor neurons in the absence and presence of SOD1 mutants. These SOD1 mutant dependent changes are likely to give insights into the pathogenesis of ALS.  

 Selected Publications 

1.      Zhu, H., Pan, S., Gu, S., Bradbury, E. M. and Chen, X. “Amino Acid Residue-Specific Stable Isotope Labeling for Quantitative Proteomics”, (2002) Rapid Communications in Mass Spectrometry, 16, 2115-2123.

2.      Zhu, H., Hunter, T. C., Pan, S., Bradbury, E. M., Chen, X. “Residue-specific mass signatures for the efficient identification of protein modifications by mass spectrometry”, (2002) Analytical Chemistry, 74, 1687-1694.

3.      Hunter, T., Yang, L., Zhu, H., Majidi, V., Bradbury, E. M., Chen, X. “Peptide mass mapping constrained with stable isotope-tagged peptides for identification of protein mixtures”, (2001) Analytical Chemistry, 73, 4891-4902.

4.      Liu, H., Zhu, H., Nersissian, A. M., Eggers, D. K., Faull, K. F., Goto, J. J., Ai, J., Sanders-Loehr, J., Gralla, E. B. and Valentine, J. S. “Copper(2+) binding to the surface residue Cys111 of His46Arg human copper-zinc superoxide dismutase, a familial amyotrophic lateral sclerosis mutant”, (2000) Biochemistry, 39, 8125-8132.

5.      Cabelli, D. E., Riley, D., Rodriguez, J., Valentine, J. S., Zhu, H. “Models of superoxide dismutase” in Biomimetic Oxidations Catalyzed by Transition Metal Complexes Meunier, B. ed., (2000) Imperial College Press, London, 461-508.

6.      Zhu, H., Shipp, E., Sanchez, R. J., Liba, A., Stein, J., Hart, J. P., Gralla, E. B., Nersissian, A. M. and Valentine, J. S. “Cobalt(2+) binding to human and tomato copper chaperone for superoxide dismutase: implications for the metal ion transfer mechanism” (2000) Biochemistry, 39, 5413-5421.

7.      Hall, L. T., Sanchez, R. J., Holloway, S. P., Zhu, H., Stine, J. E., Lyons, T. J., Demeler, B., Schirf, V., Hansen, J. C., Nersissian, A. M., Valentine, J. S., and Hart, P. J. “X-ray crystallographic and analytical ultracentrifugation analyses of truncated and full-length yeast copper chaperone for SOD (LYS7): a dimer-dimer model of Lys7-SOD association and copper delivery”, (2000) Biochemistry, 39, 3611-3623.

8.      Goto, J. J., Zhu, H., Sanchez, R. J., Nersissian, A. M., Gralla, E. B., Valentine, J. S.  and Cabelli, D. E. “Loss of in vitro metal ion binding specificity in mutant copper-zinc superoxide dismutases associated with familial amyotrophic lateral sclerosis” (2000) J. Biol. Chem., 275, 1007-1014.

9.      Zhu, H., Whitelegge, J. P., Faull, K. F., Nersissian, A. M., and Valentine, J. S. “Copper mediated oxygenation of the thiolate ligand in His46Cys mutant of yeast copper-zinc superoxide dismutase” (1999) Protein Science, 8 (Suppl. 1), 72.

10.  Lyons, T. J., Nersissian, A., Goto, J. J., Zhu, H., Gralla, E. B. and Valentine, J. S., “Metal ion reconstitution studies of yeast copper-zinc superoxide dismutase: the “phantom” subunit and the possible role of Lys7p” (1998) J. Biol. Inorg. Chem., 3, 650-662.

Group Members

Haining Zhu, P. I.

Email: haining@uky.edu 

 
Kei Fukada, postdoctoral fellow

Email: kfuda2@uky.edu

 
Fujian Zhang, graduate student

Email: fzhan2@uky.edu

Jason Garrison, rotation IBS graduate student

Email: jbgarr0@uky.edu 

Instruments and Facilities

ABI QSTAR XL quadrupole time-of-flight mass spectrometer

Protana nano-electrospray source

LC Packings nano-LC 

oMALDI source 

Amersham 2D gel electrophoresis (IPGhore and Dalt 6)

Routine molecular biology equipments for PCR, cloning, mutagenesis, tissue culture, etc.