IDENTIFICATION OF A TRANSLOCATED PROTEIN SEGMENT IN A VOLTAGE-DEPENDENT CHANNEL

被引:145
作者
SLATIN, SL
QIU, XQ
JAKES, KS
FINKELSTEIN, A
机构
[1] Albert Einstein College of Medicine, Department of Physiology and Biophysics, Bronx
关键词
D O I
10.1038/371158a0
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
VOLTAGE-GATED channels undergo a conformational change in response to changes in transmembrane voltage. Here we use site-directed biotinylation to create conformation-sensitive sites on colicin Ia, a bacteriocidal protein that forms a voltage-sensitive membrane channel, which can be monitored by electrophysiological methods(1,2). We investigated a model of gating developed for the partly homologous colicin E1 that is based on the insertion of regions of the protein into the membrane in response to cis-positive voltages(3-6). Site-directed cysteine mutagenesis, followed by chemical modification, was used to attach a biotin molecule covalently to a series of unique sites on colicin Ia. The modified protein was incorporated into planar lipid membranes, where the introduced biotin moiety served as a site to bind the water-soluble protein streptavidin, added to one side of the membrane or the other. Our results show that colicin gating is associated with the translocation across the membrane of a segment of the protein of at least 31 amino acids.
引用
收藏
页码:158 / 161
页数:4
相关论文
共 19 条
[1]   IDENTIFICATION OF A TRANSLOCATED GATING CHARGE IN A VOLTAGE-DEPENDENT CHANNEL - COLICIN-E1 CHANNELS IN PLANAR PHOSPHOLIPID-BILAYER MEMBRANES [J].
ABRAMS, CK ;
JAKES, KS ;
FINKELSTEIN, A ;
SLATIN, SL .
JOURNAL OF GENERAL PHYSIOLOGY, 1991, 98 (01) :77-93
[2]   TRANSFER OF PROTEINS ACROSS MEMBRANES .1. PRESENCE OF PROTEOLYTICALLY PROCESSED AND UNPROCESSED NASCENT IMMUNOGLOBULIN LIGHT-CHAINS ON MEMBRANE-BOUND RIBOSOMES OF MURINE MYELOMA [J].
BLOBEL, G ;
DOBBERSTEIN, B .
JOURNAL OF CELL BIOLOGY, 1975, 67 (03) :835-851
[3]   STRUCTURE AND DYNAMICS OF THE COLICIN-E1 CHANNEL [J].
CRAMER, WA ;
COHEN, FS ;
MERRILL, AR ;
SONG, HY .
MOLECULAR MICROBIOLOGY, 1990, 4 (04) :519-526
[4]  
CRONAN JE, 1990, J BIOL CHEM, V265, P10327
[5]  
DUCHE D, 1994, J BIOL CHEM, V269, P6332
[6]   THE SPONTANEOUS INSERTION OF PROTEINS INTO AND ACROSS MEMBRANES - THE HELICAL HAIRPIN HYPOTHESIS [J].
ENGELMAN, DM ;
STEITZ, TA .
CELL, 1981, 23 (02) :411-422
[7]   USE OF BIFUNCTIONAL BIOTINYL COMPOUNDS TO DETERMINE ARRANGEMENT OF SUBUNITS IN AVIDIN [J].
GREEN, NM ;
KONIECZNY, L ;
TOMS, EJ ;
VALENTINE, RC .
BIOCHEMICAL JOURNAL, 1971, 125 (03) :781-+
[8]   FLUORESCENCE ENERGY-TRANSFER DISTANCE MEASUREMENTS - THE HYDROPHOBIC HELICAL HAIRPIN OF COLICIN-A IN THE MEMBRANE-BOUND STATE [J].
LAKEY, JH ;
DUCHE, D ;
GONZALEZMANAS, JM ;
BATY, D ;
PATTUS, F .
JOURNAL OF MOLECULAR BIOLOGY, 1993, 230 (03) :1055-1067
[9]   DNA AND AMINO-ACID SEQUENCE-ANALYSIS OF STRUCTURAL AND IMMUNITY GENES OF COLICIN-IA AND COLICIN-IB [J].
MANKOVICH, JA ;
HSU, CH ;
KONISKY, J .
JOURNAL OF BACTERIOLOGY, 1986, 168 (01) :228-236
[10]   IDENTIFICATION OF A VOLTAGE-RESPONSIVE SEGMENT OF THE POTENTIAL-GATED COLICIN-E1 ION CHANNEL [J].
MERRILL, AR ;
CRAMER, WA .
BIOCHEMISTRY, 1990, 29 (37) :8529-8534