Abstract
The action of carp complement on sensitized cells (EA) proceeds in two steps requiring Ca2+and Mg2+, respectively. In order to decide which of these steps precedes in immune hemolysis, we carried out a kinetic experiment by use of a buffer and carp serum depleted of cations by passage through Amberlite IRC-50. In a reaction mixture composed of EA (sheep red blood cells sensitized with carp antibody), carp complement, optimal Ca2+and optimal Mg2+, the hemolysis took place after a lag period of about 10 min, but when Ca2+was present at zero-time and Mg2+was added 20 min later, there was almost no lag. On the contrary, when Mg2+was present at zero-time and Ca2+was added 20 min later, there was a lag of about 20 min. These observations indicate that the step requiring Ca2+precedes that in which Mg2+functions. A hemolytic intermediate complex, which was provisionally designated EACx’, was prepared by treating EA with carp complement in the presence of Ca2+, but with the absence of Mg2+at 20°C for 10 min. Furthermore, carp serum free from Cx’, which was provisionally designated Cy’, was also obtained by treating resin-passed carp serum with excessive EA in the presence of Ca2+but absence of Mg2+at 0°C for 3 h. It was confirmed that the cells in the state EACx’ were completely lysed when treated with a sufficient quantity of Cy’ and optimal Mg2+, and that Cy’ did not react with EA even in the presence of optimal Mg2+.
| Original language | English |
|---|---|
| Pages (from-to) | 841-846 |
| Number of pages | 6 |
| Journal | Nippon Suisan Gakkaishi |
| Volume | 51 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - Jan 1 1985 |
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All Science Journal Classification (ASJC) codes
- Aquatic Science
Cite this
Two Activation Steps of Carp Complement Requiring Ca2+and Mg2+and an Intermediate Product in Immune Hemolvsis. / Yano, Tomoki; Ando, Hiroko; Nakao, Miki.
In: Nippon Suisan Gakkaishi, Vol. 51, No. 5, 01.01.1985, p. 841-846.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Two Activation Steps of Carp Complement Requiring Ca2+and Mg2+and an Intermediate Product in Immune Hemolvsis
AU - Yano, Tomoki
AU - Ando, Hiroko
AU - Nakao, Miki
PY - 1985/1/1
Y1 - 1985/1/1
N2 - The action of carp complement on sensitized cells (EA) proceeds in two steps requiring Ca2+and Mg2+, respectively. In order to decide which of these steps precedes in immune hemolysis, we carried out a kinetic experiment by use of a buffer and carp serum depleted of cations by passage through Amberlite IRC-50. In a reaction mixture composed of EA (sheep red blood cells sensitized with carp antibody), carp complement, optimal Ca2+and optimal Mg2+, the hemolysis took place after a lag period of about 10 min, but when Ca2+was present at zero-time and Mg2+was added 20 min later, there was almost no lag. On the contrary, when Mg2+was present at zero-time and Ca2+was added 20 min later, there was a lag of about 20 min. These observations indicate that the step requiring Ca2+precedes that in which Mg2+functions. A hemolytic intermediate complex, which was provisionally designated EACx’, was prepared by treating EA with carp complement in the presence of Ca2+, but with the absence of Mg2+at 20°C for 10 min. Furthermore, carp serum free from Cx’, which was provisionally designated Cy’, was also obtained by treating resin-passed carp serum with excessive EA in the presence of Ca2+but absence of Mg2+at 0°C for 3 h. It was confirmed that the cells in the state EACx’ were completely lysed when treated with a sufficient quantity of Cy’ and optimal Mg2+, and that Cy’ did not react with EA even in the presence of optimal Mg2+.
AB - The action of carp complement on sensitized cells (EA) proceeds in two steps requiring Ca2+and Mg2+, respectively. In order to decide which of these steps precedes in immune hemolysis, we carried out a kinetic experiment by use of a buffer and carp serum depleted of cations by passage through Amberlite IRC-50. In a reaction mixture composed of EA (sheep red blood cells sensitized with carp antibody), carp complement, optimal Ca2+and optimal Mg2+, the hemolysis took place after a lag period of about 10 min, but when Ca2+was present at zero-time and Mg2+was added 20 min later, there was almost no lag. On the contrary, when Mg2+was present at zero-time and Ca2+was added 20 min later, there was a lag of about 20 min. These observations indicate that the step requiring Ca2+precedes that in which Mg2+functions. A hemolytic intermediate complex, which was provisionally designated EACx’, was prepared by treating EA with carp complement in the presence of Ca2+, but with the absence of Mg2+at 20°C for 10 min. Furthermore, carp serum free from Cx’, which was provisionally designated Cy’, was also obtained by treating resin-passed carp serum with excessive EA in the presence of Ca2+but absence of Mg2+at 0°C for 3 h. It was confirmed that the cells in the state EACx’ were completely lysed when treated with a sufficient quantity of Cy’ and optimal Mg2+, and that Cy’ did not react with EA even in the presence of optimal Mg2+.
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U2 - 10.2331/suisan.51.841
DO - 10.2331/suisan.51.841
M3 - Article
AN - SCOPUS:85004562291
VL - 51
SP - 841
EP - 846
JO - Nippon Suisan Gakkaishi
JF - Nippon Suisan Gakkaishi
SN - 0021-5392
IS - 5
ER -