Proteomics & Lipidomics analysis in development and disease; mass spectrometry technology development.
The Schey lab is interested in both method/instrument development in proteomics analysis as well as in applications of state-of-the-art proteomics technologies in the study of development and disease. Method development/technology projects include methods for integral membrane protein analysis, tandem mass spectrometry of intact proteins (top-down proteomics), spatially-resolved proteomics analysis, and quantitative proteomics. The major areas of application are human lens protein modifications in cataractogenesis , retina biochemical alterations in age-related macular degeneration, and antibody/antigen interactions. Other areas of interest are: exosome proteomics, Alzheimer’s disease, and aquaporin biochemistry.
Integral membrane proteins play key roles in signaling, transport, and adhesion, yet they remain difficult to analyze using conventional proteomics methods. Our lab has developed methods to analyze integral membrane proteins and membrane associated proteins including their modifications. In addition, we have developed methods for global membrane proteome analysis. Currently we are developing methods for spatially-resolved studies (see below), for tissue imaging of membrane proteins, and for examining the membrane protein-protein and protein-lipid interactions using native mass spectrometry.
Spatially-resolved proteomics represents a critically important area of development that takes advantage of recent advances in instrumental sensitivity. The ability to examine region-specific changes in a proteome has wide applicability in both development and disease processes. Multiple approaches are being developed in our lab to address this issue: 1) laser capture microsdissection (LCM) coupled with shotgun proteomics analysis, 2) MALDI imaging mass spectrometry (IMS), and 3) single cell proteomics. We have combined our membrane protein analysis protocols with LCM capture to examine region-specific changes in the ocular lens membrane proteome. Also, we have developed new sample preparation methods to allow MALDI IMS of membrane proteins. The MALDI IMS method is being applied to lens aging, diseases of the retina, and neurodegenerative diseases.
Our long-standing project on ocular lens membrane proteins is focused on how these proteins change as lens cells differentiate and as they age. Since the proteins in the lens core are as old as the individual, the lens serves as a model tissue to study protein aging. At least five mechanisms of protein-protein crosslinking have been identified. Moreover, age- and cataract-specific modifications have been identified and regional changes in the membrane proteome are being examined. Functional consequences of lens protein modification are also being examined with confocal microscopic, molecular biology, and spectroscopic methods.
The human immune system creates antibodies in response to viral threats such as SARS Coronavirus 2 (SARS-CoV-2), influenza (Flu), human immunodeficiency virus (HIV), or Ebola/Marburg viruses. The Schey lab, in collaboration with the laboratories of Drs. James Crowe and Jens Meiler, uses hydrogen-deuterium exchange (HDX) MS and crosslinking mass spectrometry (XL-MS) to elucidate antibody-antigen binding sites. With a better understanding of paratope-epitope interactions, novel broad spectrum neutralizing antibodies can be designed.