Is myelin basic protein crystallizable?
Myelin basic protein (MBP) is the predominant extrinsic protein in both central and peripheral nervous system myelins. It is thought to be involved in the stabilizing interactions between myelin membranes, and it may play an important role in demyelinating diseases such as multiple sclerosis. In spite of the fact that this abundant protein has been known for almost three decades, its three-dimensional crystal structure has not yet been determined. In this study we report on our extensive attempts to crystallize the major 18.5 kDa isoform of MBP. We used MBP having different degrees of purity, ranging from crude MBP (that was acid or salt extracted from isolated myelin), to highest purity single isoform. We used convention strategies in our search for a suitable composition of a crystallization medium. We applied both full and incomplete factorial searches for crystallization conditions. We analyzed the available data on proteins which have previously resisted crystallization, and applied this information to our own experiments. Nevertheless, despite our efforts which included 4600 different conditions, we were unable to induce crystallization of MBP. Previous work on MBP indicates that when it is removed from its native environment in the myelin membrane and put in crystallization media, the protein adopts a random coil conformation and persists as a population of structurally non-identical molecules. This thermodynamically preferred state presumably hinders crystallization, because the most fundamental factor of protein crystallization - homogeneity of tertiary structure--is lacking. We conclude that as long as its random coil flexibility is not suppressed, 18.5 kDa MBP and possibly also its isoforms will remain preeminent examples of proteins that cannot be crystallized.
Division of Neurology Research, Children's Hospital, Boston, Massachusetts.
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J
Sedzik
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DA
Kirschner
D000645:Ammonium Sulfate; D000818:Animals; D002412:Cations; D002417:Cattle; D055598:Chemical Phenomena; D011232:Chemical Precipitation; D002627:Chemistry, Physical; D003460:Crystallization; D000431:Ethanol; D005612:Freeze Drying; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances; D004676:Myelin Basic Protein / Q000737:chemistry*; D009186:Myelin Sheath / Q000737:chemistry; D011487:Protein Conformation; D011817:Rabbits; D012995:Solubility; D013816:Thermodynamics; D014961:X-Ray Diffraction
D002412:Cations; D046911:Macromolecular Substances; D004676:Myelin Basic Protein; D000431:Ethanol; D000645:Ammonium Sulfate