Purification and characterization of the assembly factor P17 of the lipid-containing bacteriophage PRD1

被引:11
作者
Caldentey, J
Hänninen, AL
Holopainen, JM
Bamford, JKH
Kinnunen, PKJ
Bamford, DH
机构
[1] Univ Helsinki, Viikki Bioctr, Inst Biotechnol, FIN-00014 Helsinki, Finland
[2] Univ Helsinki, Viikki Bioctr, Dept Biosci, FIN-00014 Helsinki, Finland
[3] Univ Helsinki, Inst Biomed, Dept Med Chem, FIN-00014 Helsinki, Finland
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1999年 / 260卷 / 02期
关键词
bacteriophage PRD1; assembly factor; macromolecular assembly; virus structure; viral protein;
D O I
10.1046/j.1432-1327.1999.00202.x
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
Assembly factors, proteins assisting the formation of viral structures, have been found in many viral systems. The gene encoding the assembly factor P17 of bacteriophage PRD1 has been cloned and expressed in Escherichia coli. P17 acts late in phage assembly, after capsid protein folding and multimerization, and sorting of membrane proteins has occurred. P17 has been purified to near homogeneity. It is a tetrameric protein displaying a rather high heat stability. The protein is largely in an cl-helical conformation and possesses a putative leucine zipper which is not essential for protein function, as judged by in vitro mutagenesis and complementation analysis. Although heating does not cause structural changes in the conformation of the protein, the dissociation of the tetramer into smaller units is evident as diminished self-quenching of the fluorescently labeled P17. Similarly, dissociation of the tetramer is also obtained by dialysis of the protein against 6-M guanidine hydrochloride (GdnHC1) or 1% SDS. The reassembly of these smaller units upon cooling is evident from resonance energy transfer.
引用
收藏
页码:549 / 558
页数:10
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