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.

Study Summary

Number of data points 332
Proteins Phosphocarrier protein HPr
Unique complexes 96
Assays/Quantities/Protocols Experimental Assay: Relative Binding Constant (Jel42) ; Experimental Assay: Relative Binding Constant (Jel44) ; Experimental Assay: Relative Binding Constant (Jel323) ; Experimental Assay: PTS (sugar phosphotransferase system) Activity ; Derived Quantity: ΔΔG (Jel323) ; Derived Quantity: ΔΔG (Jel44) ; Derived Quantity: ΔΔG (Jel42)
Libraries Binding associated with Jel42 ; Binding associated with Jel44 and Jel323; PTS activity

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
1VRC 2005-02-21T00:00:00+0000 0 Complex of enzyme IIAmannose and the histidine-containing phosphocarrier protein HPr from escherichia coli nmr, restrained regularized mean structure
2LRK 2012-04-06T00:00:00+0000 0 Solution Structures of the IIA(Chitobiose)-HPr complex of the N,N'-Diacetylchitobiose
2LRL 2012-04-06T00:00:00+0000 0 Solution Structures of the IIA(Chitobiose)-HPr complex of the N,N'-Diacetylchitobiose Branch of the Escherichia coli Phosphotransferase System
1GGR 2000-09-18T00:00:00+0000 0 COMPLEX OF ENZYME IIAGLC AND THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI NMR, RESTRAINED REGULARIZED MEAN STRUCTURE
1CM3 1999-05-13T00:00:00+0000 1.6 HIS15ASP HPR FROM E. COLI
1CM2 1999-05-13T00:00:00+0000 1.8 STRUCTURE OF HIS15ASP HPR AFTER HYDROLYSIS OF RINGED SPECIES.
2XDF 2010-04-30T00:00:00+0000 0 Solution Structure of the Enzyme I Dimer Complexed with HPr Using Residual Dipolar Couplings and Small Angle X-Ray Scattering
1J6T 2002-08-14T00:00:00+0000 0 COMPLEX OF ENZYME IIAMTL AND THE HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR FROM ESCHERICHIA COLI NMR, RESTRAINED REGULARIZED MEAN STRUCTURE
1POH 1993-10-19T00:00:00+0000 2.0 THE 2.0 ANGSTROMS RESOLUTION STRUCTURE OF ESCHERICHIA COLI HISTIDINE-CONTAINING PHOSPHOCARRIER PROTEIN HPR: A REDETERMINATION
5YA1 2017-08-29T00:00:00+0000 2.7 crystal structure of LsrK-HPr complex with ATP

Relevant UniProtKB Entries

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