| RRC ID |
15948
|
| 著者 |
Nishie M, Sasaki M, Nagao J, Zendo T, Nakayama J, Sonomoto K.
|
| タイトル |
Lantibiotic transporter requires cooperative functioning of the peptidase domain and the ATP binding domain.
|
| ジャーナル |
J Biol Chem
|
| Abstract |
Lantibiotics are ribosomally synthesized and post-translationally modified peptide antibiotics that contain unusual amino acids such as dehydro and lanthionine residues. Nukacin ISK-1 is a class II lantibiotic, whose precursor peptide (NukA) is modified by NukM to form modified NukA. ATP-binding cassette (ABC) transporter NukT is predicted to cleave off the N-terminal leader peptide of modified NukA and secrete the mature peptide. Multiple sequence alignments revealed that NukT has an N-terminal peptidase domain (PEP) and a C-terminal ATP binding domain (ABD). Previously, in vitro reconstitution of NukT has revealed that NukT peptidase activity depends on ATP hydrolysis. Here, we constructed a series of NukT mutants and investigated their transport activity in vivo and peptidase activity in vitro. Most of the mutations of the conserved residues of PEP or ABD resulted in failure of nukacin ISK-1 production and accumulation of modified NukA inside the cells. NukT(N106D) was found to be the only mutant capable of producing nukacin ISK-1. Asn(106) is conserved as Asp in other related ABC transporters. Additionally, an in vitro peptidase assay of NukT mutants demonstrated that PEP is on the cytosolic side and all of the ABD mutants as well as PEP (with the exception of NukT(N106D)) did not have peptidase activity in vitro. Taken together, these observations suggest that the leader peptide is cleaved off inside the cells before peptide secretion; both PEP and ABD are important for NukT peptidase activity, and cooperation between these two domains inside the cells is indispensable for proper functioning of NukT.
|
| 巻・号 |
286(13)
|
| ページ |
11163-9
|
| 公開日 |
2011-4-1
|
| DOI |
10.1074/jbc.M110.212704
|
| PII |
S0021-9258(20)53737-X
|
| PMID |
21303905
|
| PMC |
PMC3064170
|
| MeSH |
ATP-Binding Cassette Transporters / chemistry
ATP-Binding Cassette Transporters / genetics
ATP-Binding Cassette Transporters / metabolism*
Adenosine Triphosphate / chemistry
Adenosine Triphosphate / genetics
Adenosine Triphosphate / metabolism*
Amino Acid Substitution
Bacteriocins / chemistry
Bacteriocins / genetics
Bacteriocins / metabolism*
Gram-Positive Bacteria / enzymology*
Gram-Positive Bacteria / genetics
Mutation, Missense
Peptide Hydrolases / chemistry
Peptide Hydrolases / genetics
Peptide Hydrolases / metabolism*
Protein Binding / genetics
Protein Structure, Tertiary
|
| IF |
4.238
|
| 引用数 |
22
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| 一般微生物 |
JCM 1157 |
