TY - JOUR
T1 - Purification and properties of newt prolactin
AU - Matsuda, Kouhei
AU - Yamamoto, Kazutoshi
AU - Kikuyamai, Sakae
PY - 1990
Y1 - 1990
N2 - A highly purified prolactin (PRL) was obtained from pituitary glands of newts, Cynops pyrrhogaster, by extraction of acetone-dried powder with acid acetone and high-performance liquid chromatography (HPLC) on Mono-Q (anion exchange), Superose-12 (gel filtration), and TSK-gel ODS-120T (reverse-phase) columns with a yield of 4.5 mg per 325 mg of protein starting material. Purification was monitored by SDS-polyacrylamide gel electrophoresis (PAGE) and Western blotting analysis employing antiserum against bullfrog PRL. Newt PRL thus obtained has a molecular weight of 23,000 as determined by SDS-PAGE. The isoelectric point is 4.7 as determined by isoelectric focusing. The amino acid composition closely resembles that of anuran PRLs. This hormone was as potent as bovine PRL in stimulating collagen synthesis in the bullfrog tadpole tail fin. Antiserum against newt PRL was produced by immunizing a rabbit. Histological studies on newt adeno-hypophyses revealed that the cells that immunologically reacted with the antiserum against newt PRL correspond to the ones positively stained with the antiserum against bullfrog PRL.
AB - A highly purified prolactin (PRL) was obtained from pituitary glands of newts, Cynops pyrrhogaster, by extraction of acetone-dried powder with acid acetone and high-performance liquid chromatography (HPLC) on Mono-Q (anion exchange), Superose-12 (gel filtration), and TSK-gel ODS-120T (reverse-phase) columns with a yield of 4.5 mg per 325 mg of protein starting material. Purification was monitored by SDS-polyacrylamide gel electrophoresis (PAGE) and Western blotting analysis employing antiserum against bullfrog PRL. Newt PRL thus obtained has a molecular weight of 23,000 as determined by SDS-PAGE. The isoelectric point is 4.7 as determined by isoelectric focusing. The amino acid composition closely resembles that of anuran PRLs. This hormone was as potent as bovine PRL in stimulating collagen synthesis in the bullfrog tadpole tail fin. Antiserum against newt PRL was produced by immunizing a rabbit. Histological studies on newt adeno-hypophyses revealed that the cells that immunologically reacted with the antiserum against newt PRL correspond to the ones positively stained with the antiserum against bullfrog PRL.
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U2 - 10.1016/0016-6480(90)90206-2
DO - 10.1016/0016-6480(90)90206-2
M3 - Article
C2 - 2295424
AN - SCOPUS:0025159019
SN - 0016-6480
VL - 77
SP - 63
EP - 69
JO - General and Comparative Endocrinology
JF - General and Comparative Endocrinology
IS - 1
ER -