Year-end Sale % OFF 
Abstract
A hybrid gene consisting of the ompF promoter, the coding regions for the signal peptide, and the Ala-Glu residue of the OmpF NH2 terminus and the coding region for the major outer membrane lipoprotein devoid of the NH2-terminal cysteine residue was constructed. Escherichia coli carrying the cloned gene produced the predicted hybrid protein that is the same as the major lipoprotein except that the diacyl glycerylcysteine residue at the NH2 terminus is replaced by the Ala-Glu residue. The hybrid protein was localized in the periplasmic space as a trimer with a noncovalent interaction in addition to the previously known covalent interaction with the peptidoglycan. These results strongly indicate that the major lipoprotein exists as a trimer in the periplasmic space with covalent and noncovalent interactions with the peptidoglycan layer through the protein domain on one side and with the hydrophobic interaction with the outer membrane through the lipid domain on the other side. The trimeric structure of the lipoprotein was directly demonstrated by the chemical cross-linking of the native lipoprotein with both cleavable and uncleavable reagents. The cross-linking study also revealed interaction between the lipoprotein and the OmpA protein, a major outer membrane protein.
Highlights
IypsltainenhtlsrrncteotoeytthdrnhAsieuatgneleccalmtieyte-npidGraoeeiajnnrltorcuihdertpieinsirlocliaeidnappassutmirwdoedeepdiuidattcriehhitotci.aotsttteTetphhnidaenheattchesNopheemeaeHxyhtapcbhyzjtetrbeoriptidrriedtpmiordlrgtmpieheplrpryviaonorcitpotuowaertutnshiioenst.itiehlswneTyitdrnahhaikeisseeaapnxsltcnlooehiyasgocwrelctanleesynlsscdicaauozcesmbevlortydaas-velae-nNntahmthnoyHeaHdbtntzroetei-oiwdxxtofecinerstlgveumhtesdedenireanec,Odsahs.cmliaanoTrdpcdatooerFciimntmetpeghxarreeiaiofrnsmottNeri(dicignHneas-er,2oli“lwvftt)lheeti,hprihdismtocehfhppirhenrooyioupsmsbtssoeprisibioetndshifpi”lbpetititihridylnoOieo,ttmygewwhileypytnhhbFciccaraaoihptnrdner-ttsoadphhtsturreeesoueoiactndtcebtoiieioaawndvctteayehiassdl-e trimer in the periplasmic spawceith covalent and non- glycerylcysteine residue is replaced with the Ala-Glu residue covalentinteractions with thepeptidoglycanlayer of the OmpF NH2terminus
The biosynthetic process includes modification by glyceride andfatty acids of the precursor protein atthe cysteine residue located at the NH, terminus of the mature lipoprotein molecule [2], cleavage of the signal peptide by lipoprotein signal peptidase that islocated in the cytoplasmic membrane [3,4], acylation of the newly exposed amino group of the cysteine residue [5, 6], and translocation to the outer membrane ( 7 ) .The process has been discussed [8]
Yu et al ( 7 ) constructed a hybrid gene coding for a hybrid protein consisting of the signal peptide and the NH2-terminal 11 amino acid residues of the OmpF protein and the COOH-terminal 51 amino acid residues of the lipoprotein
Summary
IypsltainenhtlsrrncteotoeytthdrnhAsieuatgneleccalmtieyte-npidGraoeeiajnnrltorcuihdertpieinsirlocliaeidnappassutmirwdoedeepdiuidattcriehhitotci.aotsttteTetphhnidaenheattchesNopheemeaeHxyhtapcbhyzjtetrbeoriptidrriedtpmiordlrgtmpieheplrpryviaonorcitpotuowaertutnshiioenst.itiehlswneTyitdrnahhaikeisseeaapnxsltcnlooehiyasgocwrelctanleesynlsscdicaauozcesmbevlortydaas-velae-nNntahmthnoyHeaHdbtntzroetei-oiwdxxtofecinerstlgveumhtesdedenireanec,Odsahs.cmliaanoTrdpcdatooerFciimntmetpeghxarreeiaiofrnsmottNeri(dicignHneas-er,2oli“lwvftt)lheeti,hprihdismtocehfhppirhenrooyioupsmsbtssoeprisibioetndshifpi”lbpetititihridylnoOieo,ttmygewwhileypytnhhbFciccaraaoihptnrdner-ttsoadphhtsturreeesoueoiactndtcebtoiieioaawndvctteayehiassdl-e trimer in the periplasmic spawceith covalent and non- glycerylcysteine residue is replaced with the Ala-Glu residue covalentinteractions with thepeptidoglycanlayer of the OmpF NH2terminus. To construct pDOC004 carrying the ompF-lpp became sharperand clearer (lane 4 ) , suggesting thatthe hybrid gene, the 400-base pair Sau3Al fragment, that con- broadness of the bands was, at least in part, due tothe tains the lppgene lacking the region coding for the signal presence of the hybrid protein that was covalently bound to peptide and the NHz-terminal 7 amino acid residues of the the peptidoglycan fragment through the Lys residue.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
