Purification, structure and partial characterization of the major sperm activating protein complex in human serum.
Serum has long been known to enhance sperm motility, and the major factor that mediates this activation has been shown to be a macromolecule complex with a MW of about 250 kD. This macromolecule, named Sperm Activating Protein (SPAP) has now been purified by a four step protocol. Starting with 100 mL of serum 20 to 200 micrograms of purified SPAP was recovered (N = 40). Studies on the identification and structure of SPAP revealed that the macromolecule is formed by an IgG molecule to which an apolipoprotein A-I molecule is bound. Further studies on the structure of the complex was performed by partial proteolytic cleavage of SPAP by pepsin and papain and interaction with poly- and monoclonal antibodies. Based on these results the most plausible structure of SPAP places one molecule of apo A-I between the two arms of the Fab portion of an IgG4 molecule. Human spermatozoa incubated with polyclonal anti-SPAP serum and a second FITC-labelled antiserum showed fluorescence at a distinct band at the lower part of the sperm head, indicating that SPAP acts by direct interaction with the spermatozoa. The SPAP molecule shows a new mechanism mediated by the immunoglobulins, that may be very important for male fecundity.
Pousette
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A
Department of Clinical Chemistry, Karolinska Institutet, Huddinge University Hospital, Sweden.
Leijonhufvud
P K
PK
Akerlöf
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E
D016632:Apolipoprotein A-I / Q000032:analysis / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D015153:Blotting, Western; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008297:Male; D008958:Models, Molecular; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000737:chemistry* / Q000378:metabolism
D016632:Apolipoprotein A-I; D007074:Immunoglobulin G