title: Serum amyloid P component forms a stable complex with human C5b6.
abstract: A 30,000 m.w. protein bound tightly to C5b6, which was formed by activating C7-depleted human serum with zymosan. The protein remained bound to the C5b6 complex during the isolation procedure for C5b6, including chromatography on lysine-Sepharose and an anion exchange resin. Following electrophoresis and electroblotting of the C5b6 complex to a polyvinylidene difluoride transfer membrane, the 30,000 m.w. protein was microsequenced. The 24 N-terminal amino acid sequence was determined and showed identity of the 30,000 m.w. protein with the serum amyloid P (SAP) component. The C5b6-SAP complex did not dissociate in the presence of 10 mM EDTA, which distinguishes SAP-C5b6 binding from SAP's usual Ca2+-dependent binding to other molecules. SAP, which was isolated from serum by chromatography, was able to bind to preformed C5b6, which had been assembled and isolated from purified components. Functionally, the C5b6-SAP could bind C7, and the resulting C5b67-SAP complex had only moderately lower specific hemolytic activity than that of C5b67. In addition, hemolytically inactive C5b67-SAP, such as hemolytically inactive C5b67, was chemotactically active for neutrophils, while isolated SAP had no effect on cell mobility. Because SAP reacts with other serum proteins and with cells, it is likely that the addition of SAP to terminal complement complexes may affect the fate of these complexes.
authors.affiliation: Harvard-Thorndike Laboratory of the Beth Israel-Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USA.
authors.forename: S F
authors.initials: SF
authors.lastname: Barbashov
authors.forename: C
authors.initials: C
authors.lastname: Wang
authors.forename: A
authors.initials: A
authors.lastname: Nicholson-Weller
mesh_terms: D002118:Calcium / Q000378:metabolism; D003167:Complement Activation; D003165:Complement System Proteins / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D000683:Serum Amyloid P-Component / Q000378:metabolism*
chemical_list: D000683:Serum Amyloid P-Component; C050974:complement C5b-6 complex; D003165:Complement System Proteins; D002118:Calcium