Professional Education


  • Bachelor of Arts, Boston University (2014)
  • Doctor of Philosophy, Vanderbilt University (2020)
  • PhD, Vanderbilt University, Biochemistry (2020)
  • BA, Boston University, Chemistry (2014)

Stanford Advisors


All Publications


  • Glycosylation Limits Forward Trafficking of the Tetraspan Membrane Protein PMP22. The Journal of biological chemistry Marinko, J. T., Wright, M. T., Schlebach, J. P., Clowes, K. R., Heintzman, D. R., Plate, L. n., Sanders, C. R. 2021: 100719

    Abstract

    Peripheral myelin protein 22 (PMP22) folds and trafficks inefficiently, with only 20% of newly expressed protein trafficking to the cell surface. This behavior is exacerbated in many of the mutants associated with Charcot-Marie-Tooth disease (CMTD), motivating further study. Here we characterized the role of N-glycosylation in limiting PMP22 trafficking. We first eliminated N-glycosylation using an N41Q mutation, which resulted in an almost 3-fold increase in trafficking efficiency of wild type (WT) PMP22 and a 10-fold increase for the severely unstable L16P disease mutant in HEK293 cells, with similar results in Schwann cells. Total cellular levels were also much higher for the WT/N41Q mutant, though not for the L16P/N41Q form. Depletion of oligosaccharyltransferase OST-A and OST-B subunits revealed that WT PMP22 is N-glycosylated post-translationally by OST-B, whereas L16P is co-translationally glycosylated by OST-A. Quantitative proteomic screens revealed similarities and differences in the interactome for WT, glycosylation-deficient, and unstable mutant forms of PMP22 and also suggested that L16P is sequestered at earlier stages of endoplasmic reticulum (ER) quality control. CRISPR knock-out studies revealed a role for retention in ER sorting receptor 1 (RER1) in limiting the trafficking of all three forms, for UDP-glucose glycoprotein glucosyltransferase 1 (UGGT1) in limiting the trafficking of WT and L16P but not N41Q, and calnexin (CNX) in limiting the trafficking of WT and N41Q but not L16P. This work shows that N-glycosylation is a limiting factor to forward trafficking PMP22 and sheds light on the proteins involved in its quality control.

    View details for DOI 10.1016/j.jbc.2021.100719

    View details for PubMedID 33933451

  • Direct relationship between increased expression and mistrafficking of the Charcot-Marie-Tooth-associated protein PMP22. The Journal of biological chemistry Marinko, J. T., Carter, B. D., Sanders, C. R. 2020; 295 (34): 11963–70

    Abstract

    Charcot-Marie-Tooth disease (CMT) is a neuropathy of the peripheral nervous system that afflicts ∼1:2500 people. The most common form of this disease (CMT1A, 1:4000) is associated with duplication of chromosome fragment 17p11.2-12, which results in a third WT PMP22 allele. In rodent models overexpressing the PMP22 (peripheral myelin protein 22) protein and in dermal fibroblasts from patients with CMT1A, PMP22 aggregates have been observed. This suggests that overexpression of PMP22 under CMT1A conditions overwhelms the endoplasmic reticulum quality control system, leading to formation of cytotoxic aggregates. In this work, we used a single-cell flow-cytometry trafficking assay to quantitatively examine the relationship between PMP22 expression and trafficking efficiency in individual cells. We observed that as expression of WT or disease variants of PMP22 is increased, the amount of intracellular PMP22 increases to a greater extent than the amount of surface-trafficked protein. This was true for both transiently transfected cells and PMP22 stable expressing cells. Our results support the notion that overexpression of PMP22 in CMT1A leads to a disproportionate increase in misfolding and mistrafficking of PMP22, which is likely a contributor to disease pathology and progression.

    View details for DOI 10.1074/jbc.AC120.014940

    View details for PubMedID 32647009

    View details for PubMedCentralID PMC7443497

  • Peripheral myelin protein 22 preferentially partitions into ordered phase membrane domains. Proceedings of the National Academy of Sciences of the United States of America Marinko, J. T., Kenworthy, A. K., Sanders, C. R. 2020; 117 (25): 14168–77

    Abstract

    The ordered environment of cholesterol-rich membrane nanodomains is thought to exclude many transmembrane (TM) proteins. Nevertheless, some multispan helical transmembrane proteins have been proposed to partition into these environments. Here, giant plasma membrane vesicles (GPMVs) were employed to quantitatively show that the helical tetraspan peripheral myelin protein 22 (PMP22) exhibits a pronounced preference for, promotes the formation of, and stabilizes ordered membrane domains. Neither S-palmitoylation of PMP22 nor its putative cholesterol binding motifs are required for this preference. In contrast, Charcot-Marie-Tooth disease-causing mutations that disrupt the stability of PMP22 tertiary structure reduce or eliminate this preference in favor of the disordered phase. These studies demonstrate that the ordered phase preference of PMP22 derives from global structural features associated with the folded form of this protein, providing a glimpse at the structural factors that promote raft partitioning for multispan helical membrane proteins.

    View details for DOI 10.1073/pnas.2000508117

    View details for PubMedID 32513719

    View details for PubMedCentralID PMC7322011

  • Folding and Misfolding of Human Membrane Proteins in Health and Disease: From Single Molecules to Cellular Proteostasis. Chemical reviews Marinko, J. T., Huang, H. n., Penn, W. D., Capra, J. A., Schlebach, J. P., Sanders, C. R. 2019; 119 (9): 5537–5606

    Abstract

    Advances over the past 25 years have revealed much about how the structural properties of membranes and associated proteins are linked to the thermodynamics and kinetics of membrane protein (MP) folding. At the same time biochemical progress has outlined how cellular proteostasis networks mediate MP folding and manage misfolding in the cell. When combined with results from genomic sequencing, these studies have established paradigms for how MP folding and misfolding are linked to the molecular etiologies of a variety of diseases. This emerging framework has paved the way for the development of a new class of small molecule "pharmacological chaperones" that bind to and stabilize misfolded MP variants, some of which are now in clinical use. In this review, we comprehensively outline current perspectives on the folding and misfolding of integral MPs as well as the mechanisms of cellular MP quality control. Based on these perspectives, we highlight new opportunities for innovations that bridge our molecular understanding of the energetics of MP folding with the nuanced complexity of biological systems. Given the many linkages between MP misfolding and human disease, we also examine some of the exciting opportunities to leverage these advances to address emerging challenges in the development of therapeutics and precision medicine.

    View details for DOI 10.1021/acs.chemrev.8b00532

    View details for PubMedID 30608666

    View details for PubMedCentralID PMC6506414

  • Peripheral myelin protein 22 alters membrane architecture. Science advances Mittendorf, K. F., Marinko, J. T., Hampton, C. M., Ke, Z. n., Hadziselimovic, A. n., Schlebach, J. P., Law, C. L., Li, J. n., Wright, E. R., Sanders, C. R., Ohi, M. D. 2017; 3 (7): e1700220

    Abstract

    Peripheral myelin protein 22 (PMP22) is highly expressed in myelinating Schwann cells of the peripheral nervous system. PMP22 genetic alterations cause the most common forms of Charcot-Marie-Tooth disease (CMTD), which is characterized by severe dysmyelination in the peripheral nerves. However, the functions of PMP22 in Schwann cell membranes remain unclear. We demonstrate that reconstitution of purified PMP22 into lipid vesicles results in the formation of compressed and cylindrically wrapped protein-lipid vesicles that share common organizational traits with compact myelin of peripheral nerves in vivo. The formation of these myelin-like assemblies depends on the lipid-to-PMP22 ratio, as well as on the PMP22 extracellular loops. Formation of the myelin-like assemblies is disrupted by a CMTD-causing mutation. This study provides both a biochemical assay for PMP22 function and evidence that PMP22 directly contributes to membrane organization in compact myelin.

    View details for DOI 10.1126/sciadv.1700220

    View details for PubMedID 28695207

    View details for PubMedCentralID PMC5498104