Marc J. Glucksman, Ph.D. Professor & Chair, Biochemistry & Molecular Biology Director, Midwest Proteome Center Structural Neurobiology and Proteomics Laboratory PhD Columbia University, New York, NY, USA
Phone: 847.578.3235 Fax: 847.578.3240 Email: marc.glucksman@rosalindfranklin.edu
Current Research
STRUCTURAL NEUROBIOLOGY of PROCESSING ENZYMES - The degradation and/or processing of bioactive peptides is one of the most prevalent and efficient modes of physiologic regulation. My research program involves examination of structure-activity relationships at the atomic level of the enzymes and neuropeptide substrates of several reactions involved in the normal and disease states of neurobiological, aging and reproductive processes. Molecular biological methods are utilized for the generation and expression of wild type and mutagenized proteins in large quantities and cloning related enzymes while the biophysical techniques of X-ray diffraction and spectroscopy elucidate the structural detail of these macromolecules. The results of these methodologies are coupled with computer-aided modeling with homologous metalloproteases to design rational functional correlates which are then tested physiologically, and may result in better designed therapeutics. The mammalian zinc metalloendopeptidase EC 3.4.24.15 [EP 24.15], is crucial to the formation and degradation of many bioactive peptides. EP 24.15 can control mammalian reproduction through its regulation of the pivotal neuropeptide, the decapeptide gonadotropin releasing hormone [GnRH]. EP 24.15 also acts upon small peptide substrates: neurotensin, substance P, bradykinin and generating bioactive enkephalins from precursor proteins. Additional roles of this enzyme's involvement in neuropsychiatric disorders are: the ability to produce amyloidogenic peptides in Alzheimer's disease and an involvement in schizophrenia mediated by neuropeptides acting on glutamate receptors during puberty.
A. ALZHEIMER'S DISEASE - EP24.15 immunoreactivity with regions containing large pyramidal neurons in hippocampus, as well as layers 3 and 5 of the cortex. B. EP24.15 Immunoreactive in amyloid deposits and intense staining of the dystrophic neurites.
PROTEOMICS - Involves a broad-based, novel mass screening procedure to identify markers and assess structure-function relationships for diagnostics, risk factors and targets for treatment of Alzheimer's Disease, schizophrenia and endocrine cancers (e.g. prostate). New markers, notable for their diagnostic potential and prognostic values, are sought. An optimal approach is delineation of the expressed proteins for markers from human tissue and matched controls (including urine and serum) from tissue banks representing characterized patient populations. In the case of prostate cancer, initial proteomic analyses utilize cell models amenable to subcellular fractionation. We can enrich for low abundance proteins by searching components of media for secreted proteins and/or membranes for specific cell surface probes. Proteome (protein complement of the genome) paradigms commence through examining protein expression between disease and normal tissues (or cell lines). A given gene can produce more than one functional protein by: differential mRNA splicing and post-translational modifications (e.g. phosphorylation, glycosylation). Proteins are separated by 2D gel electrophoresis and changes in quantity or migration of proteins are located, quantified, excised, digested, and analyzed by mass spectroscopy. With potential "hits" from both gene and protein analyses, we utilize the approaches delineated above to identify and structurally assess proteins by X-ray crystallography as well as by defining "functional" changes present in the development and progression of diseases.
STRUCTURE-FUNCTION CORRELATES IN GENE DISCOVERY - Structure-Function Assessment and Molecular Modeling in Various Genetic Disorders: In collaboration with the Martignetti Lab at Mt. Sinai, my group utilizes computer-aided protein modeling as well as molecular biology/biochemistry and X-ray crystallographic approaches to explain the phenotype of mutations in non muscle myosin platelet disorders [May-Heggelin, Fechtner, and Sebastian; the first use of the complete chromosome 22 sequence from the Human Genome Project]. Another project involves the matrix metalloprotease 2 (MMP-2), and its involvement in a syndromic osteolysis/arthritis. This is the first proven disorder in which a matrix metalloprotease is involved and that proteolysis of extracellular matrix mediates human growth and development. In addition to molecular modeling, expression of wild type and mutant MMP-2's are evaluated for differences in their X-ray structures and kinetic characteristics towards developing therapeutics. Lastly, a putative tumour suppressor, KLF6, was assessed for potential functionality and mutants in prostate cancer assessed by the gene discovery/structure function correlate approach. Validation of these novel gene products is proceeding with proteomics and knock-in paradigms.
BOOKS
SELECTED PUBLICATIONS
(to download a PDF file of the publication, click on the reference)
Russo LC, Goñi CN, Castro LM, Asega AF, Camargo AC, Trujillo CA, Ulrich H, Glucksman MJ, Scavone C, Ferro ES. Interaction with calmodulin is important for the secretion of thimet oligopeptidase following stimulation. FEBS J. 276(16):4358-71 [2009]. medline Boudreau AC, Ferrario CR, Glucksman MJ, Wolf ME. Signaling pathway adaptations and novel protein kinase A substrates related to behavioral sensitization to cocaine. J Neurochem. 110(1):363-77 [2009]. medline Tuysuz B, Mosig R, Altun G, Sancak S, Glucksman MJ, Martignetti JA. A novel matrix metalloproteinase 2 (MMP2) terminal hemopexin domain mutation in a family with multicentric osteolysis with nodulosis and arthritis with cardiac defects. Eur J Hum Genet. 17(5):565-72 [2009]. medline Bruce LA, Sigman JA, Randall D, Rodriguez S, Song MM, Dai Y, Elmore DE, Pabon A, Glucksman MJ, Wolfson AJ. Hydrogen bond residue positioning in the 599-611 loop of thimet oligopeptidase is required for substrate selection. FEBS J. 275(22):5607-17 [2008]. medline Huang JY, Bruno AM, Patel CA, Huynh AM, Philibert KD, Glucksman MJ, Marr RA. Human membrane metallo-endopeptidase-like protein degrades both beta-amyloid 42 and beta-amyloid 40. Neuroscience. 155(1):258-62 [2008]. medline Lindvall LE, Kormeili T, Chen E, Ramirez MC, Grum-Tokars V, Glucksman MJ, Martignetti JA, Zaragoza MV, Dyson SW. Infantile systemic hyalinosis: Case report and review of the literature. J Am Acad Dermatol. 58(2):303-7 [2008]. medline Roberts JL, Mani SK, Woller MJ, Glucksman MJ, Wu TJ. LHRH-(1-5): a bioactive peptide regulating reproduction. Trends Endocrinol Metab. 18(10):386-92 [2007]. medline
Scholle MD, Kriplani U, Pabon A, Sishtla K, Glucksman MJ, Kay BK Mapping protease substrates using a biotinylated phage substrate library. Chem.Bio.Chem.7(5):834-8. [2006] medline
Wu TJ, Glucksman MJ, Roberts JL, Mani SK. Facilitation of Lordosis in Rats by a Metabolite of Luteinizing Hormone Releasing Hormone (LHRH). Endocrinology 147:2544-2549 [2006] medline
Jeske NA, Berg KA, Cousins JC, Ferro ES, Clarke WP, Glucksman MJ, Roberts JL. Modulation of bradykinin signaling by EP24.15 and EP24.16 in cultured trigeminal ganglia. J Neurochem. 97, 13-21 [2006] medline
Garcia AL, Han SK, Janssen WG, Khaing ZZ, Ito T, Glucksman MJ, Benson DL, Salton SR. A prohormone convertase cleavage site within a predicted alpha-helix mediates sorting of the neuronal and endocrine polypeptide VGF into the regulated secretory pathway. J Biol Chem. 280:41595-608 [2005] medline
Oliveira V, Garrido PA, Rodrigues CC, Colquhoun A, Castro LM, Almeida PC, Shida CS, Juliano MA, Juliano L, Camargo AC, Hyslop S, Roberts JL, Grum-Tokars V, Glucksman MJ, Ferro ES. Calcium modulates endopeptidase 24.15 membrane association, secondary structure and substrate specificity. FEBS J. 272:2978-92 [2005] medline
Sigman JA, Patwa TH, Tablante AV, Joseph CD, Glucksman MJ, Wolfson AJ. Flexibility in substrate recog-nition by thimet oligopeptidase as revealed by denaturation studies. Biochem J. 388, 255-61. [2005] medline
Wu TJ, Mani SK, Glucksman MJ, Roberts JL. Stimulatory Effect of Luteinizing Hormone-Releasing Hormone (LHRH) Metabolite, LHRH(1-5), on LHRH Gene Expression in GT1-7 Cells. Endocrinology 146, 280-6. [2005] medline
Swanson TA, Kim SI, Myers M, Pabon A, Philibert KD, Wang M, Glucksman MJ. The role of neuropeptide processing enzymes in endocrine (prostate) cancer: EC 3.4.24.15 (EP24.15) Prot Pept Lett 11, 471-78 [2004] medline
Jeske NA, Glucksman MJ, Roberts,JL. Metalloendopeptidase EC3.4.24.15 is constitutively released from the exofacial leaflet of lipid rafts in GT1-7 cells. J Neurochem. 90, 819-28 [2004] medline
Kim SI, Voshol H, van Oostrum J, Hastings TG, Cascio M, Glucksman MJ. Neuroproteomics- Expression Profiling of the Brain’s Proteomes in Health and Disease Neurochem Res 29, 1317-31 [2004] medline
Cotter EJ, Sweeney NO, Coen PM, Birney YA, Glucksman MJ, Cahill PA, Cummins PM. Regulation of endopeptidases EC3.4.24.15 and EC3.4.24.16 in vascular endothelial cells by cyclic strain: role of Gi protein signaling. Arterioscler Thromb Vasc Biol. 24, 457-63 [2004] medline
Jeske N, Glucksman MJ, Roberts J EP24.15 is Associated with Lipid Rafts J. Neurosci Res. 74, 468-73 [2003] medline
Kim SI, Grum-Tokars V, Swanson TA, Cotter EJ, Cahill PA, Roberts JL, Cummins PM Glucksman MJ Novel Roles of Neuropeptide Processing Enzymes: EC3.4.24.15 in the Neurome. J. Neurosci Res. 74, 456-67 [2003] medline
Voshol H, Glucksman MJ, van Oostrum J. Proteomics in the discovery of new therapeutic targets for psychiatric disease. Curr Mol Med. 2003 Aug;3(5):447-58. medline
Dowling O, Difeo A, Ramirez MC, Tukel T, Narla G, Bonafe L, Kayserili H, Yuksel-Apak M, Paller AS, Norton K, Teebi AS, Grum-Tokars V, Martin GS, Davis GE, Glucksman MJ, Martignetti JA. Mutations in capillary morphogenesis gene-2 result in the allelic disorders juvenile hyaline fibromatosis and infantile systemic hyalinosis. Am J Hum Genet. 2003 Oct;73(4):957-66. medline
Sigman JA, Sharky ML, Walsh ST, Pabon A, Glucksman MJ, Wolfson AJ. Involvement of surface cysteines in activity and multimer formation of thimet oligopeptidase. Protein Eng. 2003 Aug;16(8):623-8. medline
Kim SI, Pabon A, Swanson TA, Glucksman MJ. Regulation of cell-surface major histocompatibility complex class I expression by the endopeptidase EC3.4.24.15 (thimet oligopeptidase). Biochem J. 2003 Oct 1;375(Pt 1):111-20. medline
Sigman JA, Edwards SR, Pabon A, Glucksman MJ, Wolfson AJ. pH dependence studies provide insight into the structure and mechanism of thimet oligopeptidase (EC 3.4.24.15). FEBS Lett. 2003 Jun 19;545(2-3):224-8. medline
Narla G, Heath KE , Reeves HL, Li D, Giono LE , Kimmelman AC, Glucksman MJ, Narla J, Eng F, Chan AM, Ferrari A, Martignetti JA, Friedman SL KLF6, a candidate tumor suppressor gene mutated in prostate cancer. SCIENCE 2001 Dec 21; 294: 2563-2566. medline
Heath KE, Campos-Barros A, Toren A, Rozenfeld-Granot G, Carlsson LE, Savige J, Denison JC, Gregory MC, White JG, Barker DF, Greinacher A, Epstein CJ, Glucksman MJ, Martignetti JA. Nonmuscle myosin heavy chain iia mutations define a spectrum of autosomal dominant macrothrombocytopenias: may-hegglin anomaly and fechtner, sebastian, epstein, and alport-like syndromes. Am J Hum Genet. 2001 Nov;69(5):1033-45. medline
Terasawa E, Busser BW, Luchansky LL, Sherwood NM, Jennes L, Millar RP, Glucksman MJ, Roberts JL. Presence of luteinizing hormone-releasing hormone fragments in the rhesus monkey forebrain. J Comp Neurol. 2001 Oct 29;439(4):491-504. medline
Martignetti JA, Aqeel AA, Sewairi WA, Boumah CE, Kambouris M, Mayouf SA, Sheth KV, Eid WA, Dowling O, Harris J, Glucksman MJ, Bahabri S, Meyer BF, Desnick RJ. Mutation of the matrix metalloproteinase 2 gene (MMP2) causes a multicentric osteolysis and arthritis syndrome. Nat Genet. 2001 Jul;28(3):261-5. medline
Tullai J, Cummins PM, Pabon A, Roberts J, Lopingco M, Shrimpton CN, Smith AI, Martignetti J, Ferro ES, Glucksman MJ. The neuropeptide processing enzyme EC 3.4.24.15 is modulated by protein kinase A phosphorylation. J Biol Chem 2000 Nov 24;275(47):36514-22. medline
Heath,KE Babcock,M Glucksman,MJ Aliprandis,E Bizzaro,N Desnick,RJ Martignetti,JA & The May Hegglin/ Fechtner Syndrome Consortium group. Mutations in the nonmuscle myosin heavy chain IIA gene (MYH9) result in the diverse phenotypes of the May-Hegglin anomaly, Fechtner and Sebastian syndromes. Nat Genet. 2000 Sep;26(1):103-5. medline
Ferro ES, Tullai J, Glucksman MJ, Roberts J. Secretion of metalloendopeptidase 24.15 (EC 3.4.24.15). DNA Cell Biol 1999 Oct;18(10):781-9. medline
Massarelli,EE Casatti,CA Kato,A Camargo,ACM Bauer,JA Glucksman,MJ Roberts,JL Hirose,S & Ferro,ES Differential subcellular distribution of neurolysin (EC 3.4.24.16) and thimet oligopeptidase (EC 3.4.24.15) in the rat brain. Brain Res. 851, 261 [1999] medline
Cummins PM, Pabon A, Margulies EH, Glucksman MJ. Zinc coordination and substrate catalysis within the neuropeptide processing enzyme endopeptidase EC 3.4.24.15. Identification of active site histidine and glutamate residues. J Biol Chem 1999 Jun 4;274(23):16003-9. medline
Crack PJ, Wu T, Cummins PM, Ferro ES, Tullai J, Glucksman MJ, Roberts J. The association of metalloendopeptidase EC 3.4.24.15 at the extracellular surface of the AtT-20 cell plasma membrane. Brain Res 1999 Jul 24;835(2):113-24. medline
Wu T, Pierotti AR, Jakubowski M, Sheward WJ, Glucksman MJ, Smith AI, King JC, Fink G, Roberts J. Endopeptidase EC 3.4.24.15 presence in the rat median eminence and hypophysial portal blood and its modulation of the luteinizing hormone surge. J Neuroendocrinol 1997 Nov;9(11):813-22. medline
Shrimpton CN, Glucksman MJ, Lew RA, Tullai J, Margulies EH, Roberts J, Smith AI. Thiol activation of endopeptidase EC 3.4.24.15. A novel mechanism for the regulation of catalytic activity. J Biol Chem 1997 Jul 11;272(28):17395-9. medline
Meckelein B, de Silva HA, Roses AD, Rao PN, Pettenati MJ, Xu PT, Hodge R, Glucksman MJ, Abraham CR. Human endopeptidase (THOP1) is localized on chromosome 19 within the linkage region for the late-onset alzheimer disease AD2 locus. Genomics 1996 Jan 15;31(2):246-9. medline
Course Information:
Medical School: MBCH 505A, B & C Medical Biochemistry MBCH 502 Molecular Cell Biology GIGP 501 Molecular Cell Biology Graduate School: BC 5254/GCS 719 Computer Applications in Biomedical Research GBCH 543 Enzyme Structure and Mechanism GBCH 532 Biochemistry and Molecular Biology Journal Club GBCH 535 Doctoral Dissertation in Biochemistry
Last updated by Correll November 2, 2010