Combinatorial peptide libraries reveal the ligand-binding mechanism of the oligopeptide receptor OppA of Lactococcus lactis

被引:65
作者
Detmers, FJM
Lanfermeijer, FC
Abele, R
Jack, RW
Tampé, R
Konings, WN
Poolman, B
机构
[1] Univ Groningen, Groningen Biomol Sci & Biotechnol Inst, Dept Biochem, NL-9747 AG Groningen, Netherlands
[2] Univ Groningen, Groningen Biomol Sci & Biotechnol Inst, Dept Microbiol, NL-9747 AG Groningen, Netherlands
[3] Univ Marburg, Inst Physiol Chem, D-35033 Marburg, Germany
[4] EMC Microcollect, D-72070 Tubingen, Germany
关键词
D O I
10.1073/pnas.220308797
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
The oligopeptide transport system (Opp) of Lactococcus lactis has the unique capacity to mediate the transport of peptides from 4 up to at least 18 residues. The substrate specificity of this binding protein-dependent ATP-binding cassette transporter is determined mainly by the receptor protein OppA. To study the specificity and ligand-binding mechanism of OppA, the following strategy was used: (i) OppA was purified and anchored via the lipid moiety to the surface of liposomes; (ii) the proteoliposomes were used in a rapid filtration-based binding assay with radiolabeled nonameric bradykinin as a reporter peptide; and (iii) combinatorial peptide libraries were used to determine the specificity and selectivity of OppA. The studies show that (i) OppA is able to bind peptides up to at least 35 residues, but there is a clear optimum in affinity for nonameric peptides; (ii) the specificity for nonameric peptides is not equally distributed over the whole peptide, because positions 4, 5. and 6 in the binding site are more selective; and (iii) the differences in affinity for given side chains is relatively small, but overall hydrophobic residues are favored-whereas glycine, proline, and negatively charged residues lower the binding affinity. The data indicate that not only the first six residues (enclosed by the protein) but also the C-terminal three residues interact in a nonopportunistic: manner with (the surface of) OppA, This binding mechanism is different from the one generally accepted for receptors of ATP-binding cassette-transporter systems.
引用
收藏
页码:12487 / 12492
页数:6
相关论文
共 22 条
[1]   PARTIAL-PURIFICATION OF THE 5-HYDROXYTRYPTOPHAN-REUPTAKE SYSTEM FROM HUMAN BLOOD-PLATELETS USING A CITALOPRAM-DERIVED AFFINITY RESIN [J].
BIESSEN, EAL ;
HORN, AS ;
ROBILLARD, GT .
BIOCHEMISTRY, 1990, 29 (13) :3349-3354
[2]   Relating structure to thermodynamics: The crystal structures and binding affinity of eight OppA-peptide complexes [J].
Davies, TG ;
Hubbard, RE ;
Tame, JRH .
PROTEIN SCIENCE, 1999, 8 (07) :1432-1444
[3]   Kinetics and specificity of peptide uptake by the oligopeptide transport system of Lactococcus lactis [J].
Detmers, FJM ;
Kunji, ERS ;
Lanfermeijer, FC ;
Poolman, B ;
Konings, WN .
BIOCHEMISTRY, 1998, 37 (47) :16671-16679
[4]   TRANSPORT OF BETA-CASEIN-DERIVED PEPTIDES BY THE OLIGOPEPTIDE TRANSPORT-SYSTEM IS A CRUCIAL STEP IN THE PROTEOLYTIC PATHWAY OF LACTOCOCCUS-LACTIS [J].
KUNJI, ERS ;
HAGTING, A ;
DEVRIES, CJ ;
JUILLARD, V ;
HAANDRIKMAN, AJ ;
POOLMAN, B ;
KONINGS, WN .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (04) :1569-1574
[5]   Reconstruction of the proteolytic pathway for use of β-casein by Lactococcus lactis [J].
Kunji, ERS ;
Fang, G ;
Jeronimus-Stratingh, CM ;
Bruins, AP ;
Poolman, B ;
Konings, WN .
MOLECULAR MICROBIOLOGY, 1998, 27 (06) :1107-1118
[6]   On the binding mechanism of the peptide receptor of the oligopeptide transport system of Lactococcus lactis [J].
Lanfermeijer, FC ;
Detmers, FJM ;
Konings, WN ;
Poolman, B .
EMBO JOURNAL, 2000, 19 (14) :3649-3656
[7]   Kinetics and consequences of binding of nona- and dodecapeptides to the oligopeptide binding protein (OppA) of Lactococcus lactis [J].
Lanfermeijer, FC ;
Picon, A ;
Konings, WN ;
Poolman, B .
BIOCHEMISTRY, 1999, 38 (44) :14440-14450
[8]  
LOWRY OH, 1951, J BIOL CHEM, V193, P265
[9]   ELECTROSPRAY MASS-SPECTROMETRY AND TANDEM MASS-SPECTROMETRY OF SYNTHETIC MULTICOMPONENT PEPTIDE MIXTURES - DETERMINATION OF COMPOSITION AND PURITY [J].
METZGER, JW ;
KEMPTER, C ;
WIESMULLER, KH ;
JUNG, G .
ANALYTICAL BIOCHEMISTRY, 1994, 219 (02) :261-277
[10]   Molecular recognition templates of peptides: Driving force for molecular evolution of peptide transporters [J].
Payne, JW ;
Grail, BM ;
Marshall, NJ .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2000, 267 (01) :283-289