Mink growth hormone structural-functional relationships:: Effects of renaturing and storage conditions

被引:9
作者
Borromeo, Vitaliano [1 ,2 ]
Sereikaite, Jolanta [3 ]
Bumelis, Vladas-Algirdas [3 ]
Secchi, Camillo [2 ]
Scire, Andrea [4 ]
Ausili, Alessio [4 ]
D'Auria, Sabato [5 ]
Tanfani, Fabio [4 ]
机构
[1] Univ Milan, Dept Vet Pathol Hyg & Hlth, Biochem & Physiol Unit, I-20133 Milan, Italy
[2] Univ Milan, Dept Anim Pathol & Hlth, Biochem & Physiol Unit, Fac Vet Med, I-20133 Milan, Italy
[3] Vilnius Gediminas Tech Univ, Fac Fundamental Sci, Dept Chem & Bioengn, Vilnius, Lithuania
[4] Univ Politecn Marche, Fac Sci, Inst Biochem, Ancona, Italy
[5] CNR, Inst Prot Biochem, I-80125 Naples, Italy
关键词
mink growth hormone; Fourier-transform infrared spectroscopy; biological activity; immunological activity; freeze-drying;
D O I
10.1007/s10930-007-9120-1
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Fourier-transform infrared spectroscopy, in vitro bioassay and enzyme-linked immunoassay were used to study the structural-functional relationships of recombinant mink growth hormone (mGH), refolded and stored under different conditions. Porcine GH (pGH) was synthesized and used as an example. These two hormones, when refolded and stored the same way, had the same secondary structures, biological and immunological efficacy, and biological potency. Only the immunological potency differed, mGH being significantly less potent than pGH. Renaturation pH and storing frozen or at 4 degrees C in 5% glycerol did not affect either the secondary structure or the activity. However, freeze-drying raised the content of buried alpha-helices and lowered that of solvated alpha-helices and of unordered structures. These conformational changes were associated with a reduction of immunological and biological potency of mGH and of immunological potency of pGH. These findings provide original information on the secondary structure of mGH, and show that conformational changes induced by lyophilization adversely affect its activity.
引用
收藏
页码:170 / 180
页数:11
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