Hew C L, Wang N C, Yan S, Cai H, Sclater A, Fletcher G L
Eur J Biochem. 1986 Oct 15;160(2):267-72. doi: 10.1111/j.1432-1033.1986.tb09966.x.
The precursor proteins for winter flounder antifreeze polypeptide (AFP) were isolated from liver using gel filtration chromatography and reverse-phase high-performance liquid chromatography. Two major pro-antifreezes (Mr 5000), corresponding to the precursors for AFP-6 and AFP-8, were characterized by amino acid analyses and automated Edman degradation. These precursors showed significant antifreeze activity. The pro-antifreezes were synthesized in the liver seasonally as demonstrated by immunoblotting and in vitro liver incorporation studies. No mature AFP were detected in liver, thus indicating that the processing of pro-antifreezes, including amidation of the C-termini, occurred mainly in the serum. The function(s) of the prosequences, if any, remain unclear.
利用凝胶过滤色谱法和反相高效液相色谱法从肝脏中分离出冬鲽抗冻多肽（AFP）的前体蛋白。通过氨基酸分析和自动埃德曼降解法对两种主要的前抗冻蛋白（分子量5000）进行了表征，它们分别对应于AFP-6和AFP-8的前体。这些前体表现出显著的抗冻活性。免疫印迹和体外肝脏掺入研究表明，前抗冻蛋白在肝脏中季节性合成。在肝脏中未检测到成熟的AFP，因此表明前抗冻蛋白的加工，包括C末端的酰胺化，主要发生在血清中。前导序列的功能（如果有的话）仍不清楚。
