Creatine kinase-mediated ATP supply fuels actin-based events in phagocytosis.
Phagocytosis requires locally coordinated cytoskeletal rearrangements driven by actin polymerization and myosin motor activity. How this actomyosin dynamics is dependent upon systems that provide access to ATP at phagosome microdomains has not been determined. We analyzed the role of brain-type creatine kinase (CK-B), an enzyme involved in high-energy phosphoryl transfer. We demonstrate that endogenous CK-B in macrophages is mobilized from the cytosolic pool and coaccumulates with F-actin at nascent phagosomes. Live cell imaging with XFP-tagged CK-B and beta-actin revealed the transient and specific nature of this partitioning process. Overexpression of a catalytic dead CK-B or CK-specific cyclocreatine inhibition caused a significant reduction of actin accumulation in the phagocytic cup area, and reduced complement receptor-mediated, but not Fc-gammaR-mediated, ingestion capacity of macrophages. Finally, we found that inhibition of CK-B affected phagocytosis already at the stage of particle adhesion, most likely via effects on actin polymerization behavior. We propose that CK-B activity in macrophages contributes to complement-induced F-actin assembly events in early phagocytosis by providing local ATP supply.
Kuiper
Jan W P
JW
Department of Cell Biology, Radboud University, Nijmegen Medical Centre, Nijmegen, The Netherlands.
Pluk
Helma
H
Oerlemans
Frank
F
van Leeuwen
Frank N
FN
de Lange
Frank
F
Fransen
Jack
J
Wieringa
Bé
B
D000199:Actins / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism* / Q000502:physiology* / Q000600:supply & distribution; D000818:Animals; D002448:Cell Adhesion; D003165:Complement System Proteins / Q000378:metabolism; D052276:Creatine Kinase, BB Form / Q000378:metabolism* / Q000502:physiology; D003404:Creatinine / Q000031:analogs & derivatives / Q000494:pharmacology; D017737:Macrophages, Peritoneal / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D050505:Mutant Proteins / Q000378:metabolism; D009895:Opsonin Proteins / Q000378:metabolism; D010587:Phagocytosis / Q000502:physiology; D010588:Phagosomes / Q000378:metabolism; D011108:Polymers / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology; D013997:Time Factors; D015054:Zymosan / Q000378:metabolism
D000199:Actins; D050505:Mutant Proteins; D009895:Opsonin Proteins; D011108:Polymers; C012260:cyclocreatine; D000255:Adenosine Triphosphate; D003165:Complement System Proteins; D015054:Zymosan; D003404:Creatinine; D052276:Creatine Kinase, BB Form