Formation of high-affinity C5 convertases of the alternative pathway of complement.
Cleavage of C5 by C5 convertase is the last enzymatic step in the complement activation cascade leading to the formation of the cytolytic proteolytically activated form of C5 (C5b)-9 complex. In the present study, we examined the effect of the density of C3b (the proteolytically activated form of C3) on the function of the noncatalytic subunit of natural surface-bound forms of the enzyme. A comparison of the kinetic parameters of C5 convertases assembled on three surfaces (zymosan, rabbit erythrocytes, and sheep erythrocytes) were similar and revealed that the average K:(m) decreased approximately 28-fold (5.2-0.18 microM) when the density of C3b was increased from approximately 18,000 to 400,000 C3b/cell. Very-high-affinity C5 convertases were generated when preformed C3 convertases were allowed to self amplify by giving them excess C3. These convertases exhibited K(m) from 0.016 to 0.074 microM, well below the normal plasma concentration of C5 in blood (0.37 microM). The results suggest that in serum convertases formed with monomeric C3b will be relatively inefficient in capturing C5 but will continue to cleave C3 opsonizing the cell surface for phagocytosis, whereas convertases formed with C3b-C3b complexes in areas of high C3b density will primarily cleave C5. The catalytic rate of these convertases approaches maximum velocity, thereby switching the enzyme from cleavage of C3 to cleavage of C5, and production of the cytolytic C5b-9 complex.
Rawal
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N
Department of Biochemistry, University of Texas Health Science Center, Tyler, TX 75708, USA. nenoo.rawal@uthct.edu
Pangburn
M
M
D000818:Animals; D001665:Binding Sites / Q000276:immunology; D002384:Catalysis; D002645:Chickens; D050577:Complement C3-C5 Convertases / Q000097:blood / Q000378:metabolism*; D003179:Complement C3b / Q000378:metabolism; D003182:Complement C5 / Q000378:metabolism; D015941:Complement Hemolytic Activity Assay; D003170:Complement Pathway, Alternative / Q000276:immunology; D004910:Erythrocyte Membrane / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D007700:Kinetics; D011485:Protein Binding / Q000276:immunology; D011817:Rabbits; D012756:Sheep; D015054:Zymosan / Q000276:immunology / Q000378:metabolism
D003182:Complement C5; D003179:Complement C3b; D015054:Zymosan; D050577:Complement C3-C5 Convertases