Epitope mapping by mutagenesis distinguishes between the two tertiary structures of the histidine-containing protein HPr.


Abstract

Thirty-four of the 85 residues of the histidine-containing protein HPr of the Escherichia coli phosphoenolpyruvate:sugar phosphotransferase system have been changed by site-directed mutagenesis. Many of the mutations have wild-type activity suggesting an unaltered tertiary structure but have altered binding to three monoclonal antibodies: Jel42, Jel44, and Jel323. This altered binding defines the residues that are involved in the epitopes of HPr. At present, two different three-dimensional structures have been determined for HPr, one from two-dimensional nuclear magnetic resonance spectra and the other from x-ray diffraction of HPr crystals. The epitope mapping for Jel42 does not distinguish between the tertiary structures. However, only the HPr structure derived from two-dimensional nuclear magnetic resonance spectra is consistent with a contiguous surface binding site that can be defined as the epitope for Jel44. Thus the x-ray structure may represent a partially unfolded HPr.

Submission Details

ID: xawcnBDF

Submitter: Shu-Ching Ou

Submission Date: Feb. 27, 2019, 12:36 p.m.

Version: 1

Publication Details
Sharma S;Georges F;Delbaere LT;Lee JS;Klevit RE;Waygood EB,Proc Natl Acad Sci U S A (1991) Epitope mapping by mutagenesis distinguishes between the two tertiary structures of the histidine-containing protein HPr. PMID:1711212
Additional Information

Sequences for Jel44 and Jel323 are not found, thus the sequences/mutants associated with the binding data do not include sequences of these two monoclonal antibodies.

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1PFH 1995-11-14 THE PHOSPHORYLATED FORM OF THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR
1VRC 2005-04-19 Complex of enzyme IIAmannose and the histidine-containing phosphocarrier protein HPr from escherichia coli nmr, restrained regularized mean structure
1HDN 1994-06-22 THE HIGH-RESOLUTION STRUCTURE OF THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI DETERMINED BY RESTRAINED MOLECULAR DYNAMICS FROM NMR NUCLEAR OVERHAUSER EFFECT DATA
3EZE 1998-12-16 COMPLEX OF THE AMINO TERMINAL DOMAIN OF ENZYME I AND THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI NMR, RESTRAINED REGULARIZED MEAN STRUCTURE
1GGR 2000-11-15 COMPLEX OF ENZYME IIAGLC AND THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI NMR, RESTRAINED REGULARIZED MEAN STRUCTURE
3EZB 1999-12-16 COMPLEX OF THE AMINO TERMINAL DOMAIN OF ENZYME I AND THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI
2LRL 2012-05-16 Solution Structures of the IIA(Chitobiose)-HPr complex of the N,N'-Diacetylchitobiose Branch of the Escherichia coli Phosphotransferase System
3EZA 1999-05-25 COMPLEX OF THE AMINO TERMINAL DOMAIN OF ENZYME I AND THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI NMR, RESTRAINED REGULARIZED MEAN STRUCTURE
2LRK 2012-05-16 Solution Structures of the IIA(Chitobiose)-HPr complex of the N,N'-Diacetylchitobiose
2XDF 2010-09-22 Solution Structure of the Enzyme I Dimer Complexed with HPr Using Residual Dipolar Couplings and Small Angle X-Ray Scattering
1J6T 2002-11-13 COMPLEX OF ENZYME IIAMTL AND THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI NMR, RESTRAINED REGULARIZED MEAN STRUCTURE
3CCD 2008-10-21 1.0 1.0 A Structure of Post-Succinimide His15Asp HPr
1OPD 1997-08-20 1.5 HISTIDINE-CONTAINING PROTEIN (HPR), MUTANT WITH SER 46 REPLACED BY ASP (S46D)
1CM3 2000-05-17 1.6 HIS15ASP HPR FROM E. COLI
1CM2 2000-05-17 1.8 STRUCTURE OF HIS15ASP HPR AFTER HYDROLYSIS OF RINGED SPECIES.
1POH 1994-01-31 2.0 THE 2.0 ANGSTROMS RESOLUTION STRUCTURE OF ESCHERICHIA COLI HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR: A REDETERMINATION
4XWJ 2015-10-21 2.1 Histidine-containing phosphocarrier protein (HPr) and antisigma factor Rsd complex
2JEL 1998-05-27 2.5 JEL42 FAB/HPR COMPLEX
5YA2 2018-07-11 2.7 Crystal structure of LsrK-HPr complex with ADP
5YA1 2018-07-11 2.7 crystal structure of LsrK-HPr complex with ATP
5YA0 2018-07-11 3.0 Crystal structure of LsrK and HPr complex
6KCR 2019-10-09 3.5 X-ray structure of the MtlR-HPr complex from Escherichia coli

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Phosphocarrier protein HPr P0AA06 PTHP_ECO57
100.0 Phosphocarrier protein HPr P0AA05 PTHP_ECOL6
100.0 Phosphocarrier protein HPr P0AA04 PTHP_ECOLI
100.0 Phosphocarrier protein HPr P0AA08 PTHP_SALTI
100.0 Phosphocarrier protein HPr P0AA07 PTHP_SALTY
100.0 Phosphocarrier protein HPr P0AA09 PTHP_SHIFL
98.8 Phosphocarrier protein HPr P16481 PTHP_KLEPN