A hydroxyproline-containing protein from shark brain that is related to myelin basic protein.
Myelin basic protein (MBP) from shark (Chondricthyes) consists of a simpler mixture of charge isomers than human MBP. About two-thirds of the total amount applied to a CM-52 cellulose cation-exchange column was recovered in the unbound fraction of the column; the remaining one-third bound to column and was eluted as a single OD280 peak. This bound material did not sow the usual pattern of charge microheterogeneity found with human or bovine MBP. The unbound fraction was composed of a high molecular weight protein (55-60 kDa), which constituted most of this protein fraction and a low molecular weight protein (approximately 18 kDa). The amino acid composition of our unbound fraction was similar to that reported earlier. The Glx (glutamic acid + glutamine) was increased about threefold whereas the Arg content was only about 25% of that of the 18.5 kDa variant of bovine or human origin. The presence of hydroxyproline (1.2 residues/100) in this protein was noteworthy, identification of which was achieved by amino acid analysis in two different systems and by mass spectrometry. In the precolumn derivatization method, hydroxyproline eluted at 2.7 min; in the postcolumn derivatization method it eluted at 12.2 min. Identification of hydroxyproline was completed by fast atom bombardment-mass spectral analysis. The effect of hydroxyproline on the secondary structure of this protein is being studied. Verification that this high molecular weight protein contained MBP sequences within its primary structure was confirmed by immunological methods.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Biochemistry, Hospital for Sick Children, Toronto, Ontario, Canada.
D D
DD
Wood
J
J
McLaurin
M A
MA
Moscarello
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D006909:Hydroxyproline / Q000378:metabolism*; D004676:Myelin Basic Protein / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D012754:Sharks
D000596:Amino Acids; D004676:Myelin Basic Protein; D009419:Nerve Tissue Proteins; D006909:Hydroxyproline