Sequence-related behaviour of transmembrane domains from class I receptor tyrosine kinases

被引:16
作者
Jones, DH [1 ]
Barber, KR [1 ]
Grant, CWM [1 ]
机构
[1] Univ Western Ontario, Dept Biochem, London, ON N6A 5C1, Canada
来源
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES | 1998年 / 1371卷 / 02期
基金
英国医学研究理事会;
关键词
deuterium NMR; peptide; model membrane; signal transduction; cholesterol; freeze-fracture;
D O I
10.1016/S0005-2736(98)00015-7
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
H-2 NMR spectroscopy and freeze-fracture elect:ron microscopy were used to compare the transmembrane domains of two Class I protein receptor tyrosine kinases (the EGF receptor and Neu/erbB-2) regarding overall behaviour in fluid lipid bilayer membranes. The 34-residue peptide, EGFR(tm), was synthesised to contain the 23 amino acid hydrophobic stretch (Ile(622) to Met(644)) thought to span the membrane of the human EGF receptor, plus the first 10 amino acids (Arg(645) to Thr(654)) of the cytoplasmic domain. Deuterium probes replaced selected H-1 nuclei at sites corresponding to Ala(623), Met(644), and Val(650). The 38-residue peptide, Neu(tm), was synthesised having the 21 residue hydrophobic stretch (Ile(660) to Ile(680)) calculated to span the membrane in rat Neu/erbB-2, plus residues Lys(681) to Thr(691) of the contiguous cytoplasmic domain. Deuterium probes replaced selected H-1 nuclei at Ala(661), Leu(667), and Val(676). A third peptide, Neu(tm)*, was also prepared, corresponding to the transmembrane domain of a constitutively-activating Neu/erbB-2 transformant in which Val(664) is replaced by Glu: it was deuterated in a manner identical to Neu(tm). Peptides were studied by H-2 NMR spectroscopy at 1 mol% and 6 mol% in unsonicated fluid bilayers of 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) and in POPC containing 33 mol% cholesterol, over the range 12 degrees to 65 degrees C. Overall motion was found to be different for each of the three peptides under a given set of conditions. EGFR(tm) spectra were characteristic of axially symmetric motion in membranes of POPC alone, and in POPC/cholesterol at 35 degrees C and above. In contrast, spectra of the transmembrane peptides, Neu(tm) and Neu(tm) *, were characteristic of significantly axially asymmetric motion under all conditions studied (and regardless of sample preparation method). Addition of 33% cholesterol to membranes was accompanied by spectral changes consistent with increased formation of peptide dimers/oligomers in all cases. The transformant peptide, Neu(tm) *, showed greater spectral evidence of immobilisation than did the wild type - probably reflecting a greater tendency to form large oligomers. Sequence-related details within the transmembrane domains of Class I receptor tyrosine kinases appear to exert important control over their associations within membranes. Freeze-fracture electron microscopy of the NMR samples demonstrated their liposomal nature. Peptide-related intramembranous particles (IMPss) were present which likely represent oligomers of the transmembrane peptide. IMP size and distribution were similar under a given set of conditions for all three peptides, suggesting that the differences seen by NMR spectroscopy reflect structures smaller than the 2 nm resolution limit of freeze-fracture EM and peptide relationships within its 20 nm accuracy of identifying lateral position. (C) 1998 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:199 / 212
页数:14
相关论文
共 81 条
[1]   Ligand-independent dimerization of oncogenic v-erbB products involves covalent interactions [J].
Adelsman, MA ;
Huntley, BK ;
Maihle, NJ .
JOURNAL OF VIROLOGY, 1996, 70 (04) :2533-2544
[2]  
[Anonymous], METHOD ENZYMOL
[3]   ELUCIDATION OF MOTIONAL MODES IN GLYCOGLYCEROLIPID BILAYERS - A H-2 NMR RELAXATION AND LINE-SHAPE STUDY [J].
AUGER, M ;
CARRIER, D ;
SMITH, ICP ;
JARRELL, HC .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1990, 112 (04) :1373-1381
[4]   ONCOGENIC ACTIVATION OF THE NEU-ENCODED RECEPTOR PROTEIN BY POINT MUTATION AND DELETION [J].
BARGMANN, CI ;
WEINBERG, RA .
EMBO JOURNAL, 1988, 7 (07) :2043-2052
[5]   DEUTERIUM QUADRUPOLE ECHO NMR-STUDY OF METHYL-GROUP DYNAMICS IN N-ACETYL-DL-(GAMMA-D6)-VALINE [J].
BESHAH, K ;
GRIFFIN, RG .
JOURNAL OF MAGNETIC RESONANCE, 1989, 84 (02) :268-274
[6]   INTRAPEPTIDE AUTOPHOSPHORYLATION OF THE EPIDERMAL GROWTH-FACTOR RECEPTOR - REGULATION OF KINASE CATALYTIC FUNCTION BY RECEPTOR DIMERIZATION [J].
BISWAS, R ;
BASU, M ;
SENMAJUMDAR, A ;
DAS, M .
BIOCHEMISTRY, 1985, 24 (14) :3795-3802
[7]   INTRAMEMBRANE HELIX-HELIX ASSOCIATION IN OLIGOMERIZATION AND TRANSMEMBRANE SIGNALING [J].
BORMANN, BJ ;
ENGELMAN, DM .
ANNUAL REVIEW OF BIOPHYSICS AND BIOMOLECULAR STRUCTURE, 1992, 21 :223-242
[8]   High-speed magic angle spinning solid-state H-1 nuclear magnetic resonance study of the conformation of gramicidin a in lipid bilayers [J].
Bouchard, M ;
Davis, JH ;
Auger, M .
BIOPHYSICAL JOURNAL, 1995, 69 (05) :1933-1938
[9]   EVIDENCE FOR SIMILAR FUNCTION OF TRANSMEMBRANE SEGMENTS IN RECEPTOR AND MEMBRANE-ANCHORED PROTEINS [J].
BRANDL, CJ ;
DEBER, RB ;
HSU, LC ;
WOOLLEY, GA ;
YOUNG, XK ;
DEBER, CM .
BIOPOLYMERS, 1988, 27 (07) :1171-1182
[10]   CONFORMATIONAL-CHANGES INDUCED BY THE TRANSFORMING AMINO-ACID SUBSTITUTION IN THE TRANSMEMBRANE DOMAIN OF THE NEU ONCOGENE-ENCODED P185 PROTEIN [J].
BRANDTRAUF, PW ;
PINCUS, MR ;
CHEN, JM .
JOURNAL OF PROTEIN CHEMISTRY, 1989, 8 (06) :749-756