Plasminogen binds specifically to alpha-enolase on rat neuronal plasma membrane.
Plasminogen (PGn) that we identified in microglial-conditioned medium has a neurotrophic factor-like effect on cultured neurons. We have also shown that PGn binds specifically to a protein with a molecular mass of 45 kDa in the neuronal plasma membrane. As a candidate PGn receptor-like molecule on the neuronal surface, this 45-kDa protein was purified from the plasma membrane of embryonic rat brain. Amino acid sequence analysis of polypeptides derived from the cleavage of the protein with cyanogen bromide and V8 protease revealed that the 45-kDa protein is identical to rat alpha-enolase. In fact, PGn was found to bind to purified rat alpha-enolase and also to a synthetic peptide (30 residues) that corresponds to the carboxyl terminal region of rat alpha-enolase. Physical properties of the 45-kDa protein, such as molecular mass, isoelectric point, and the ability to form dimers, are quite similar to those of alpha-enolase. The 45-kDa PGn-binding protein in the plasma membrane was also recognized by anti-rat alpha-enolase antibody, and pretreatment with alpha-enolase antibody markedly diminished the PGn-binding to the plasma membrane. In addition, immunocytochemical staining of the cultured cells under the nonpermeable condition showed that alpha-enolase is present on the cell surface of a certain population of neurons. These results suggest that alpha-enolase may function as a PGn-binding molecule on the neuronal cell surface.
Department of Neurochemistry, National Institute of Neuroscience, Tokyo.
K
K
Nakajima
M
M
Hamanoue
N
N
Takemoto
T
T
Hattori
K
K
Kato
S
S
Kohsaka
D000595:Amino Acid Sequence; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology*; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D003488:Cyanogen Bromide; D007150:Immunohistochemistry; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009474:Neurons / Q000201:enzymology*; D010446:Peptide Fragments / Q000737:chemistry; D010751:Phosphopyruvate Hydratase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010958:Plasminogen / Q000378:metabolism*; D051381:Rats; D017421:Sequence Analysis; D012697:Serine Endopeptidases
D017077:Culture Media, Conditioned; D010446:Peptide Fragments; D010958:Plasminogen; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D010751:Phosphopyruvate Hydratase; D003488:Cyanogen Bromide