My research lab is mainly focused on the development of advanced mass spectrometric methods and strategies for analysis of biological molecules such as proteins, metabolites and environmental toxins. Our current research activities include the following major projects:

1.Analysis of protein posttranslational modifications, in particular, fatty acylation and relevant phosphorylation. The major difference between prokaryotic and eukaryotic cells is the highly developed endomembrane system that divides the cell into different compartments with different membrane structures and functions. I am interested in the mechanisms how proteins are transported to different destinations across different membranes after synthesized in ribosome.The covalent attachment of lipids to proteins (Protein Posttranslational fatty acylation) plays important roles in the regulation of protein signaling, trafficking, localizing and enzymatic activities in cells and tissues. It has been found that fatty acylation not only occurs on numerous membrane proteins and membrane associated cytoplasmic proteins but also is related with phosphorylation.Phosphorylation within polybasic motifs introduces negative charge and functions as electrostatic switches.Because of the importance of protein fatty acylation in microbial pathogenesis and progression of many human diseases,my research has been working on development of mass spectrometry-based approaches to understand the molecular mechanisms of the cycles of acylation-deacylation and identify chemical inhibitors that can treat microbial infections and other human diseases.

Additionally, we have also worked on the multidimensional separation of histone variants and identification of posttranslational modifications. It is aimed to explore the epigenetic effects of environmental exposures to pesticides or other toxins.

Selected publications:

(1)Hongying Zhong*, Xiao Xiao, Shi Zheng, Wenyang Zhang, Mengjie Ding, Haiying Jiang, Lulu Huang, Jie Kang. Mass Spectrometric Analysis of Mono- and Multi- Phosphopeptides by Selective Binding with NiZnFe2O4 magnetic Nanoparticles. Nat. Commun. 2013, 4, 1656.

(2)Hongying Zhong*，Guangfu Yang*. Structure and Function Analysis of Protein Posttranslational Fatty Acylation: Targeting Lipid Signals in Cells. Science China (Chemistry) 2013，43, 35-48 (Invited). (in Chinese)

(3)Shi Zheng, Xiaoli Wang, Jieying Fu, Xuejiao Hu, Xiao Xiao, Lulu Huang, Youe Zhou, Hongying Zhong*. Desalting of Phosphopeptides by Tandem Polypyrrole-C18 Reverse Phase Micropipette Tip (TMTipPPY-C18) Based on Hybrid Electrostatic, Π-Π Stacking and Hydrophobic Interactions for Mass Spectrometric Analysis. Anal. Chim. Acta 2012,724, 73-79.

(4)Linjie Dong, Jianjian Li, Lun Li, Tingting Li, Hongying Zhong*. Comparative Analysis of S-Fatty Acylation of Gel-Separated Proteins by Stable Isotope-Coded Fatty Acid Transmethylation and Mass Spectrometry. Nat. Protoc. 2011, 6, 1377-1390.

(5)Lun Li, Linjie Dong, Lisha Xia, Tingting Li, Hongying Zhong*. Chemical and Genetic Probes for Analysis of Protein Palmitoylation. J. Chromatogr. B 2011, 879, 1316-1324. Invited by Professor Jack Rosenfeld, Department of Pathology, McMaster University.

(6)Jianjian Li, Yingxia Yue, Xuejiao Hu, Hongying Zhong*. Rapid Transmethylation and Stable Isotope Labeling for Comparative Analysis of Fatty Acids by Mass Spectrometry. Anal. Chem. 2009, 81, 5080-5087.

 2.Quantification of interactions between proteins and endogenous free fatty acids or exogenous chemicals. Blood is the key biological sample that determines the ADME (Absorption, Distribution, Metabolism, Excretion) process of exogenous drugs or toxins. Crystallographic studies can show the structure of proteins in solid state but the three dimensional structure of proteins and the interactions with small molecules in solution remain largely unknown.  We are interested in developing mass spectrometry-based approaches for quantitative assessment of the intrinsic binding affinity, the affinitive variations induced by exogenous chemicals, and the topological dependence of protein affinity.  The strategies are either based on the equilibrium partition or ion mobility.

Selected publications:

(1)Tingting Li, Yingxia Yue, Jianjian Li, Xiaoli Wang, Jieying Fu, Hongying Zhong*.  Quantification of Interactions between Serum Albumin and Endogenous Free Fatty Acids or Exogenous Chemicals by Stable Isotope-Coded Mass Spectrometry.  ACS Med. Chem. Lett. 2011, 2, 587-591.

(2)Youe Zhou, Qian Xia, Mengjie Ding, Hubert Kageruka, Haiying Jiang, Yan Jiang, Huifen Jin, Xiao Xiao, Hongying Zhong*. Magnetic Nanoparticles of Nitrogen Enriched Carbon (mnNEC) for Analysis of Pesticides and Metabolites in Zebrafish by Gas Chromatography-Mass Spectrometry. J. Chromatogr. B 2013, 915-916, 46-51.

(3)Yingxia Yue, Lun, Li, Yong Liang, Linjie Dong, Qingjian Dong, Hongying Zhong*.  Effects of Co-Existed Proteins on Measurement of Pesticide Residues in Blood by Gas Chromatography-Mass Spectrometry.J. Chromatogr. B. 2010, 878, 3089-3094.

3.Large scale data analysis and bioinformatics. Proteome and metabolome represent the complete collection of all proteins or metabolites in cells, tissues and organs. The complexity of samples results in huge data sets.  How to find useful bioinformation from massive data is the key to systems biology.  Based on the quantitative analysis of fatty acids, we have developed a hierarchical clustering approach for rapid identification of unknown microorganisms.  In the resultant dendrogram,the Euclidean distance between unknown species and known species quantitatively reveals their differences or shared similarities in fatty acid related pathways.

De novo protein sequencing based on tandem mass spectrometry is also important to identify proteins that can not be identified through database searching due to amino mutations, chemical modifications and in-complete genome databases.  Development of computational or experimental approaches is necessary to ensure accurate identification of protein sequences.

Selected publications:

(1)Tingting Li, Ling Dai, Lun Li, Xuejiao Hu, Linjie Dong, JianjianLi, Sule Khalfan Salim, Jieying Fu, Hongying Zhong*. Typing of Unknown Microorganisms Based on Quantitative Analysis of Fatty Acids by Mass Spectrometry and Hierarchical Clustering. Anal. Chim. Acta 2011, 684，8-16. Featured Article.

(2)Hongying Zhong*, Linjie Dong, Qingjian Dong, Changshu Ke, Jieying Fu, Xiaoli Wang, Cong Liu, Ling Dai. Quantitative Analysis of Aberrant Fatty Acid Composition of Zebrafish Hepatic Lipids Induced by Organochlorine Pesticides Using Stable Isotope-Coded Transmethylation and Gas Chromatography-Mass Spectrometry.  Anal. Bioanal. Chem. 2012, 404, 207-216.

4.Biomarker discovery. Because pesticides have been extensively used worldwide for agricultural purposes, a major public health concern has been raised.  Aberrant hepatic lipid composition has been a hallmark of many liver diseases associated with exposures to various chemicals and environmental toxins.  We have been working on the development of analytical 4.techniques that can quantitatively determine the changes in fatty acid composition of hepatic lipids.  In addition, quantitative measurement of changes in liver protein expression is also pursued.

Selected publications:

Hongying Zhong*, Linjie Dong, Qingjian Dong, Changshu Ke, Jieying Fu, Xiaoli Wang, Cong Liu, Ling Dai. Quantitative Analysis of Aberrant Fatty Acid Composition of Zebrafish Hepatic Lipids Induced by Organochlorine Pesticides Using Stable Isotope-Coded Transmethylation and Gas Chromatography-Mass Spectrometry.  Anal. Bioanal. Chem. 2012, 404, 207-216.

5.Development of techniques including new ionization methods based on laser activated electron tunneling (LAET, Anal. Chim. Acta 2012), stable isotope-coded derivatization, new MALDI matrix, MALDI imaging that can enable mass spectrometric analysis of biological systems.  Data acquisition and processing have also been studied.

Selected publications:

(1)Lulu Huang, Xiao Xiao, Yinping Xie, Hubert Kageruka, Youe Zhou, Fengjiao Deng, Hongying Zhong*.  Compressed matrix thin film (CMTF)-assisted laser desorption ionization mass spectrometric analysis.Anal. Chim. Acta 2013, 786, 85-94.

(2)Hongying Zhong*, Jieying Fu, Xiaoli Wang, Shi Zheng.  Measurement of Laser Activated Electron Tunneling from Semiconductor Zinc Oxide to Adsorbed Organic Molecules by a Matrix Assisted Laser Desorption Ionization Mass Spectrometer.  Anal. Chim. Acta 2012, 729, 45-53.

(3)Hongying Zhong*, Xiangzhu Xiao, Peibiao Li. Application of Fourier Transform to Four-Electrode Microvoltammetric System for Studies on Micro-ITIES Electroanalysis. Anal. Chim. Acta 1999, 382, 137-144.

Grant Awards as PI:

1.National Natural Science Foundation of China (NSFC, 31270876).

2.National Natural Science Foundation of China (NSFC, 21175054).

3.Hubei Natural Science Foundation Council (HBNSFC, 2009CDA001).

4.Research Funds of Central China Normal University from the Ministry of Education (120002040270).

5.Research Funds of Central China Normal University from the Ministry of Education (CCNU11G01007).

6.National Natural Science Foundation of China (NSFC, 21575046).

7.National Natural Science Foundation of China (NSFC, 21545401).

8.National Natural Science Foundation of China (NSFC, 31370815).

Grant Awards as Co-PI:

1.Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT, no. IRT0953).

2.The Research Platform of Hubei Province for Monitoring of Pesticide Residues and Agricultural Products Safety.

3.Research Funds of Central China Normal University from the Ministry of Education (CCNU11C01002).