TY - JOUR
T1 - Covalent binding of [35S]cysteine to the labile binding site of the third component of complement
T2 - A physiological approach
AU - Rutherford, Cynthia J.
AU - Jenkins, David R.
N1 - Funding Information:
This research was supported by a grant from the Blood Foundation
PY - 1985/10
Y1 - 1985/10
N2 - The alternative pathway of complement was activated physiologically by agarose beads to which [35S]cysteine had been bound by a disulfide link. The activated form of the third component of complement, C3b, which had bound to the radioactive cysteine was then released from tharose bead with dithiothreitol. The [35S]cysteine was shown to be covalently bound to the α′ chain of C3b, and to its known major breakdown product, the 66,000-Da polypeptide. This is highly suggestive that complement activation has led to formation of a peptide bond between the radioactive cysteine and the labile binding site on the α′ chain of C3b. This radioactive marker will enable the amino acid sequence of the labile binding site to be determined, with the knowledge that the labeling of the amino acid(s) has occurred during physiological activation.
AB - The alternative pathway of complement was activated physiologically by agarose beads to which [35S]cysteine had been bound by a disulfide link. The activated form of the third component of complement, C3b, which had bound to the radioactive cysteine was then released from tharose bead with dithiothreitol. The [35S]cysteine was shown to be covalently bound to the α′ chain of C3b, and to its known major breakdown product, the 66,000-Da polypeptide. This is highly suggestive that complement activation has led to formation of a peptide bond between the radioactive cysteine and the labile binding site on the α′ chain of C3b. This radioactive marker will enable the amino acid sequence of the labile binding site to be determined, with the knowledge that the labeling of the amino acid(s) has occurred during physiological activation.
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U2 - 10.1016/0090-1229(85)90137-0
DO - 10.1016/0090-1229(85)90137-0
M3 - Article
C2 - 4028522
AN - SCOPUS:0022273805
SN - 0090-1229
VL - 37
SP - 77
EP - 82
JO - Clinical Immunology and Immunopathology
JF - Clinical Immunology and Immunopathology
IS - 1
ER -