The N-terminal portion of the preToc75 transit peptide interacts with membrane lipids and inhibits binding and import of precursor proteins into isolated chloroplasts

被引:30
作者
Inoue, K
Demel, R
de Kruijff, B
Keegstra, K [1 ]
机构
[1] Michigan State Univ, Dept Energy, Plant Res Lab, E Lansing, MI 48824 USA
[2] Univ Utrecht, Ctr Biomembranes & Lipid Enzymol, Biomembrane Inst, Utrecht, Netherlands
[3] Michigan State Univ, Dept Biochem & Mol Biol, Dept Plant Biol, E Lansing, MI 48824 USA
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 2001年 / 268卷 / 14期
关键词
chloroplast protein import; IMPACT system; peptide-lipid interaction; Toc75; transit peptide;
D O I
10.1046/j.1432-1327.2001.02316.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
Toc75 is an outer envelope membrane protein of chloroplasts. It is unusual among the outer membrane proteins in that its precursor form has a bipartite transit peptide. The N-terminal portion of the Toc75 transit peptide is sufficient to target the protein to the stromal space of chloroplasts. We prepared a 45 amino-acid peptide containing the stromal targeting domain of the Toc75 transit peptide in Escherichia coli, using the intein-mediated system, and purified it by reverse-phase HPLC. Its identity was confirmed by N-terminal amino-acid sequencing and matrix assisted laser desorption ionization mass spectrometry. In monolayer experiments, the peptide inserted into the chloroplastic membrane lipids sulfoquinovosyl diacylglycerol and phosphatidylglycerol and into a nonchloroplastic lipid phosphatidylethanolamine. However, it did not insert into other chloroplastic lipids, such as mono- and digalactosyl diacylglycerol, and phosphatidylcholine. Furthermore, the peptide significantly inhibited binding of radiolabeled precursors of Toc75 and the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase to intact chloroplasts as effectively as did a bacterially produced precursor or the small subunit of 1,5-bisphosphate carboxylase/oxygenase. The peptide also inhibited import of radiolabeled precursors into isolated chloroplasts, however, to a lesser extent than did nonlabeled precursor or the small subunit of 1,5-bisphosphate carboxylase/oxygenase.
引用
收藏
页码:4036 / 4043
页数:8
相关论文
共 31 条
[1]   Chloroplast transit peptides: structure, function and evolution [J].
Bruce, BD .
TRENDS IN CELL BIOLOGY, 2000, 10 (10) :440-447
[2]  
Bruce BD, 1994, PLANT MOL BIOL MAN J, VJ1, P1
[3]   Single-column purification of free recombinant proteins using a self-cleavable affinity tag derived from a protein splicing element [J].
Chong, SR ;
Mersha, FB ;
Comb, DG ;
Scott, ME ;
Landry, D ;
Vence, LM ;
Perler, FB ;
Benner, J ;
Kucera, RB ;
Hirvonen, CA ;
Pelletier, JJ ;
Paulus, H ;
Xu, MQ .
GENE, 1997, 192 (02) :271-281
[4]   Import and routing of nucleus-encoded chloroplast proteins [J].
Cline, K ;
Henry, R .
ANNUAL REVIEW OF CELL AND DEVELOPMENTAL BIOLOGY, 1996, 12 :1-26
[5]   The C terminus of a chloroplast precursor modulates its interaction with the translocation apparatus and PIRAC [J].
Dabney-Smith, C ;
van den Wijngaard, PWJ ;
Treece, Y ;
Vredenberg, WJ ;
Bruce, BD .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (45) :32351-32359
[6]   Reconstitution of a chloroplast protein import channel [J].
Hinnah, SC ;
Hill, K ;
Wagner, R ;
Schlicher, T ;
Soll, J .
EMBO JOURNAL, 1997, 16 (24) :7351-7360
[7]   EXTENT OF N-TERMINAL METHIONINE EXCISION FROM ESCHERICHIA-COLI PROTEINS IS GOVERNED BY THE SIDE-CHAIN LENGTH OF THE PENULTIMATE AMINO-ACID [J].
HIREL, PH ;
SCHMITTER, JM ;
DESSEN, P ;
FAYAT, G ;
BLANQUET, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (21) :8247-8251
[8]   Identification of a Hsp70 recognition domain within the rubisco small subunit transit peptide [J].
Ivey, RA ;
Subramanian, C ;
Bruce, BD .
PLANT PHYSIOLOGY, 2000, 122 (04) :1289-1299
[9]  
Ivey RA, 2000, CELL STRESS CHAPERON, V5, P62, DOI 10.1379/1466-1268(2000)005<0062:IVAIVI>2.0.CO
[10]  
2