Expression of recombinant forms of human 21.5 kDa myelin basic protein and proteolipid protein in CHO cells

• Autorzy: Jaskiewicz E. , Jedynak A. , Ziolo E.
• Języki publikacji: EN

Abstrakty

EN

MBP and PLP are major structural protein components of myelin. Both proteins play a functional role in formation of myelin sheath and in maintenance of its compaction. Immune responses to MBP and PLP have been implicated in the pathogenesis of multiple sclerosis (MS), an auto-immune disease of the central nervous system. Recombinant forms of both proteins isolated and purified from bacterial or insect cell systems are commonly used to study the specificity of auto-response in MS. We have prepared recombinant forms of MBP and PLP stably expressed in CHO cells. Several clones with proper cytoplasmic MBP or surface PLP localization were obtained and characterized by flow cytometry and indirect immunostaining. CHO cells expressing the recombinant forms of MBP and PLP can be very useful in studies on the autoimmune mechanism of MS.

Słowa kluczowe

EN

myelin basic protein; pathogenesis; cell; multiple sclerosis; Chinese hamster ovary; expression; structural protein component; central nervous system; proteolipid protein; human disease; man; myelin; immune response

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2005
• Tom: 52
• Numer: 4
• Opis fizyczny: p.863-866,fig.,ref.

Twórcy

autor

Jaskiewicz E.

• Polish Academy of Sciences, Wroclaw, Poland

autor

Jedynak A.

autor

Ziolo E.

Bibliografia

• Bates IR, Harauz G (2003) Molecular dynamics exposes α- helices in myelin basic protein. J Mol Model 9: 290–297.
• Bates IR, Matharu P, Noboru I, Rochon D, Wood DD, Polverini E, Moscarello MA, Viner N, Harauz G (2000) Characterization of recombinant murine 18.5 kDa myelin basic protein. Protein Expr Purif 20: 285–299.
• Bates IR, Boggs JM, Feix JB, Harauz G (2003) Membraneanchoring and charge effects in the interaction of myelin basic protein with lipid bilayers studied by site-directed spin labeling. J Biol Chem 278: 9041–9047.
• Bates IR, Boggs JM, Feix JB, Harauz G (2004) An immunodominant epitope of myelin basic protein is an amphipathic alpha-helix. J Biol Chem 279: 5757–5764.
• Boison D, Stoffel W (1994) Disruption of the compacted myelin sheath of axons of the central nervous system in proteolipid protein-deficient mice. Proc Natl Acad Sci USA 91: 11709–11713.
• Bongarzone ER, Jacobs E, Schonmann V, Kampf K, Campagnoni CW, Campagnoni AT (2001) Differential sensitivity in the survival of oligodendrocyte cell lines to overexpression of myelin proteolipid protein gene products. J Neurosci Res 65: 485–492.
• Diehl HJ, Schaich M, Budzinski RM, Stoffel W (1986) Individual exons encode the integral membrane domains of human myelin proteolipid protein. Proc Natl Acad Sci USA 83: 9807–9811.
• Fukuzono S, Takeshita T, Sakamoto T, Hisada A, Shimizu N, Mikoshiba K (1998) Overproduction and immuno-affinity purification of myelin proteolipid protein (PLP), an inositol hexakisphosphate-binding protein, in a baculovirus expression system. Biochem Biophys Res Commun 249: 66–72.
• Jaskiewicz E (2004) Epitopes on myelin proteins recognized by autoantibodies present in multiple sclerosis patients. Post Hig Med Dośw 58: 472–482 (in Polish).
• Kronquist KE, Crandall BF, Macklin WB, Campagnoni AT (1987) Expression of myelin proteins in the developing human spinal cord: cloning and sequencing of human proteolipid protein cDNA. Neurosci Res 18: 395–401.
• Nave KA, Lai C, Bloom FE, Milner RJ (1987) Splice site selection in the proteolipid protein (PLP) gene transcript and primary structure of the DM-20 protein of central nervous system myelin. Proc Natl Acad Sci USA 84: 5665–5669.
• Nye SH, Pelfrey CM, Burkwit JJ, Voskuhl R, Lenardo MJ, Mueller JP (1995) Purification of immunologically active recombinant 21.5 kDa isoform of human myelin basic protein. Mol Immunol 32: 1131–1141.
• O’Conor KC, Bar-Or A, Hafler DA (2001) The neuroinmmunology of multiple sclerosis: possible roles of T and B lymphocytes in immunopathogenesis. J Clin Immunol 21: 81–92.
• Ridsdale R, Beniac DR, Tompkins T, Moscarello MA, Harauz G (1997a) Three-dimensional structure of Myelin basic protein. Reconstruction via angular reconstruction of randomly oriented single particles. J Biol Chem 272: 4261–4268.
• Ridsdale R, Beniac DR, Tompkins T, Moscarello MA, Harauz G (1997b) Three-dimensional structure of myelin basic protein. Molecular modeling and considerations of predicted structures in multiple sclerosis. J Biol Chem 272: 4269–4275.
• Roth HJ, Kronquist KE, Kerlero de Rosbo N, Crandall BF, Campagnoni AT (1987) Evidence for the expression of four myelin basic protein variants in the developing human spinal cord through cDNA cloning. J Neurosci Res 17: 321–328.
• Sedzik J, Blaurock AE, Hochli M (1984) Lipid\myelin basic protein multilayers. A model for the cytoplasmic space in central nervous system myelin. J Mol Biol 174: 385–409.
• Schmidt S (1999) Candidate autoantigens in multiple sclerosis. Mult Scler 5: 147–160.
• Stoffel W, Hollen H, Giersiefen H (1984) Structure and molecular arrangement of proteolipid protein of central nervous system myelin. Proc Natl Acad Sci USA 81: 5012–5016.
• Weimbs T, Stoffel W (1992) Proteolipid protein (PLP) of CNS myelin: positions of free, disulfide-bonded and fatty acid thioester-linked cysteine residues and implications for the membrane topology of PLP. Biochemisty 31: 12289–12296.
• Wood DD, Moscarello MA (1989) The isolation, characterization, and lipid-aggregating properties of citrulline containing myelin basic protein. J Biol Chem 264: 5121–5127.