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Purification of androgen‐binding protein from rat testis using high‐performance liquid chromatography and physicochemical properties of the iodinated molecule
Author(s) -
Gueant Jean-Louis,
Khanfri Jamal,
Gerard Hubert,
Fremont Sophie,
Gerard Annie,
Grig Georges,
Nicolas Jean-Pierre
Publication year - 1986
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(86)81505-8
Subject(s) - chemistry , chromatography , size exclusion chromatography , isoelectric focusing , ultracentrifuge , elution , molecular mass , sedimentation coefficient , high performance liquid chromatography , yield (engineering) , isoelectric point , gel permeation chromatography , biochemistry , enzyme , materials science , organic chemistry , metallurgy , polymer
The androgen‐binding protein (ABP) has been purified 87 500‐fold from rat testis using 4 steps of HPLC, with a yield of 14%. The molecule was 99% pure with a specific activity estimated to 16 600 pmol/mg protein. The iodinated molecule was eluted in 2 peaks in Sephacryl S300 gel filtration with a molecular mass estimated to be 92 600 ± 3300 and 50 300 ± 4000 Da. The column isoelectrofocusing of 125 I‐ABP demonstrated 3 isoproteins isoelectric at pH 4.7, 4.9 and 5.3 and the sedimentation coefficient was estimated to be 4.7 S in sucrose gradient ultracentrifugation. The 125 I‐ABP had similar physicochemical properties to the non‐labelled ABP of epididymis.