Identification and purification of apolipoprotein C-III from the serum of cows

Midori Yamamoto From the Fukushima Kenchu Livestock Hygiene Service Center, Kooriyama, Fukushima 963-8041 (Yamamoto); the Department of Systematic Diagnosis, National Institute of Animal Health, 3-1-1 Kannondai, Tsukuba, Ibaraki 305-0856 (Katoh); the Ibaraki University School of Agriculture, Ami, Ibaraki 300-0393 (Adachi); and the Department of Veterinary Internal Medicine, Rakuno Gakuen University, Ebetsu, Hokkaido 069-8501 (Oiknwa), Japan.

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Norio Katoh From the Fukushima Kenchu Livestock Hygiene Service Center, Kooriyama, Fukushima 963-8041 (Yamamoto); the Department of Systematic Diagnosis, National Institute of Animal Health, 3-1-1 Kannondai, Tsukuba, Ibaraki 305-0856 (Katoh); the Ibaraki University School of Agriculture, Ami, Ibaraki 300-0393 (Adachi); and the Department of Veterinary Internal Medicine, Rakuno Gakuen University, Ebetsu, Hokkaido 069-8501 (Oiknwa), Japan.

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Yoshikazu Adachi From the Fukushima Kenchu Livestock Hygiene Service Center, Kooriyama, Fukushima 963-8041 (Yamamoto); the Department of Systematic Diagnosis, National Institute of Animal Health, 3-1-1 Kannondai, Tsukuba, Ibaraki 305-0856 (Katoh); the Ibaraki University School of Agriculture, Ami, Ibaraki 300-0393 (Adachi); and the Department of Veterinary Internal Medicine, Rakuno Gakuen University, Ebetsu, Hokkaido 069-8501 (Oiknwa), Japan.

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Shin Oikawa From the Fukushima Kenchu Livestock Hygiene Service Center, Kooriyama, Fukushima 963-8041 (Yamamoto); the Department of Systematic Diagnosis, National Institute of Animal Health, 3-1-1 Kannondai, Tsukuba, Ibaraki 305-0856 (Katoh); the Ibaraki University School of Agriculture, Ami, Ibaraki 300-0393 (Adachi); and the Department of Veterinary Internal Medicine, Rakuno Gakuen University, Ebetsu, Hokkaido 069-8501 (Oiknwa), Japan.

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Abstract

Objective

To identify, purify, and analyze distribution of apolipoprotein C-III in lipoprotein fractions and to evaluate its concentration in serum from calves, heifers, and cows during various stages of lactation.

Sample Population

Sera from 3 female calves, 3 heifers, and 12 cows during early, middle, late, and nonlactation stages.

Procedure

Apolipoprotein C-III was identified by use of amino-terminal amino acid sequence analysis of bands separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Purification was performed by extraction with acetone, delipidation with 2-propanol, and 2 chromatographic steps. The apolipoprotein C-III concentration in the total lipoprotein fraction was evaluated by densitometric scanning of the bands for apolipoprotein C-III separated by electrophoresis, using purified apolipoprotein C-III as an internal standard.

Results

Apolipoprotein C-III was identified as 8.2- and 7.3-kd proteins with identical amino acid sequences. The 2 proteins were mainly found in the high-density lipoprotein fraction, then were purified separately. Serum apolipoprotein C-III concentration was significantly (P< 0.05) higher in cows during lactation than in nonlactating cows and was negatively correlated with serum triglyceride concentration (r = −0.620, P < 0.01).

Conclusions

The role of apolipoprotein C-III in cows may involve a function related to lactation. (Am J Vet Res 1998;59:667-672)

Abstract

Objective

To identify, purify, and analyze distribution of apolipoprotein C-III in lipoprotein fractions and to evaluate its concentration in serum from calves, heifers, and cows during various stages of lactation.

Sample Population

Sera from 3 female calves, 3 heifers, and 12 cows during early, middle, late, and nonlactation stages.

Procedure

Apolipoprotein C-III was identified by use of amino-terminal amino acid sequence analysis of bands separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Purification was performed by extraction with acetone, delipidation with 2-propanol, and 2 chromatographic steps. The apolipoprotein C-III concentration in the total lipoprotein fraction was evaluated by densitometric scanning of the bands for apolipoprotein C-III separated by electrophoresis, using purified apolipoprotein C-III as an internal standard.

Results

Apolipoprotein C-III was identified as 8.2- and 7.3-kd proteins with identical amino acid sequences. The 2 proteins were mainly found in the high-density lipoprotein fraction, then were purified separately. Serum apolipoprotein C-III concentration was significantly (P< 0.05) higher in cows during lactation than in nonlactating cows and was negatively correlated with serum triglyceride concentration (r = −0.620, P < 0.01).

Conclusions

The role of apolipoprotein C-III in cows may involve a function related to lactation. (Am J Vet Res 1998;59:667-672)

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