MS characterization of qualitative protein polymorphisms in the spinal cords of inbred mouse strains.
The spinal cord proteomes of two inbred mouse strains with different susceptibility to experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis, were investigated by 2-DE and MALDI-MS. A proteome map comprising 304 different protein species was established. Using 2-D fluorescence difference gel electrophoresis, a comparison of the mouse strains revealed 26 qualitatively polymorphic proteins with altered electrophoretic mobility. MS analyses and DNA sequencing were applied to characterize their structural differences and 14 single amino acid substitutions were identified. Moreover, analysis of selectively enriched phosphopeptides from the neurofilament heavy polypeptide of both mouse strains revealed a high degree of diversity in the phosphorylated C-terminal domains of this protein. The described approach is capable to structurally characterize qualitative protein polymorphisms, whereas their functional significance remains to be elucidated. For some proteins formerly associated with experimental autoimmune encephalomyelitis and/or multiple sclerosis structural polymorphisms are described here, which may be subjected to further investigations. In addition, this work should be of general interest for proteomic analysis of inbred strains, because it shows potentials and constraints in the use of 2-DE analysis and MALDI-MS to detect and characterize structural protein polymorphisms.
Mikkat
Stefan
S
Core Facility Proteome Analysis, Medical Faculty, University of Rostock, Rostock, Germany. stefan.mikkat@med.uni-rostock.de
Lorenz
Peter
P
Scharf
Christian
C
Yu
Xinhua
X
Glocker
Michael O
MO
Ibrahim
Saleh M
SM
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000235:genetics; D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016900:Neurofilament Proteins / Q000032:analysis / Q000737:chemistry; D010748:Phosphopeptides / Q000032:analysis / Q000737:chemistry; D011110:Polymorphism, Genetic; D011506:Proteins / Q000235:genetics*; D013116:Spinal Cord / Q000378:metabolism*
D000596:Amino Acids; D016900:Neurofilament Proteins; D010748:Phosphopeptides; D011506:Proteins; C051774:neurofilament protein H