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Journal of Biochemistry Advance Access originally published online on May 31, 2008
Journal of Biochemistry 2008 144(3):371-381; doi:10.1093/jb/mvn077
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© 2008 The Japanese Biochemical Society

Agonistic and Antagonistic Effects of C5a-Chimera Bearing S19 Ribosomal Protein Tail Portion on the C5a Receptor of Monocytes and Neutrophils, Respectively

Yuuichiro Oda1,2, Kazutaka Tokita1, Yoshihiko Ota1, Ying Li1, Keisuke Taniguchi1,3, Norikazu Nishino4, Katsumasa Takagi2, Tetsuro Yamamoto1 and Hiroshi Nishiura1,*

1Department of Molecular Pathology; 2Department of Orthopedic and Neuro-Musculoskeletal Surgery, Faculty of Medical and Pharmaceutical Sciences, Kumamoto University; 3Division of Pharmacology, Kumamoto Laboratory, Mitsubishi Chemical Safety Institute of Laboratory, Kumamoto; and 4Department of Biological Functions and Engineering, Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology, Kitakyushu, Japan

*To whom correspondence should be addressed. Tel: 81963735306, Fax: 81963735308, E-mail: seino{at}kumamoto-u.ac.jp

Received May 11, 2008; Accepted May 24, 2008


  Abstract

C-terminus of S19 ribosomal protein (RP S19) endows the cross-linked homodimer with a dual effect on the C5a receptor in leucocyte chemoattraction; agonistic effect on the monocyte receptor, and antagonistic effect on the neutrophil receptor. C5a exhibits the uniform agonistic effect on this receptor of both cell types. We have currently prepared a recombinant C5a-chimeric protein bearing the C-terminus of RP S19 (C5a/RP S19 chimera) to be used as a substitute of the RP S19 dimer. In vitro, this chimera similarly inhibited the intracellular Ca2+ mobilization of neutrophils induced by C5a to the RP S19 dimer did. In the guinea pig skin, 10–7 M C5a/RP S19 chimera exhibited an inhibitory capacity to the neutrophil infiltration induced by 3 x 10–7 M C5a without enhancing monocyte infiltration. In reverse passive Arthus reaction, the neutrophil infiltration associated with plasma extravasation was significantly reduced by the simultaneous administration of 10–7 M C5a/RP S19 chimera with antibodies. The C5a/RP S19 chimera is a useful tool not only to examine the molecular mechanism that underlies the functional difference of the C5a receptor between monocytes and neutrophils, but also to prevent C5a-mediated hyper-response of neutrophils in acute inflammation.

Key Words: C5a, C5a receptor, monocytes, neutrophils, S19 ribosomal protein

Abbreviations: BSA, bovine serum albumin; C5a/RP S19 chimera, recombinant C5a-chimeric protein bearing the C-terminus of RP S19; FBS, fetal bovine serum; fMLP, formyl-Met-Leu-Phe; HBSS, Hanks’ balanced salt solution; HPLC, high performance liquid column chromatography; OVA, ovalbumin; PAGE, polyacrylamide gel electrophoresis; RP S19, ribosomal protein S19; SDS, sodium dodecyl sulphate


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