Thermodynamic effects of replacements of Pro residues in helix interiors of maltose-binding protein.


Abstract

Introduction of Pro residues into helix interiors results in protein destabilization. It is currently unclear if the converse substitution (i.e., replacement of Pro residues that naturally occur in helix interiors would be stabilizing). Maltose-binding protein is a large 370-amino acid protein that contains 21 Pro residues. Of these, three nonconserved residues (P48, P133, and P159) occur at helix interiors. Each of the residues was replaced with Ala and Ser. Stabilities were characterized by differential scanning calorimetry (DSC) as a function of pH and by isothermal urea denaturation studies as a function of temperature. The P48S and P48A mutants were found to be marginally more stable than the wild-type protein. In the pH range of 5-9, there is an average increase in T(m) values of P48A and P48S of 0.4 degrees C and 0.2 degrees C, respectively, relative to the wild-type protein. The other mutants are less stable than the wild type. Analysis of the effects of such Pro substitutions in MBP and in three other proteins studied to date suggests that substitutions are more likely to be stabilizing if the carbonyl group i-3 or i-4 to the mutation site is not hydrogen bonded in the wild-type protein. Study holds ProTherm entries: 17404, 17405, 17406, 17407, 17408, 17409, 17410, 17411, 17412, 17413, 17414, 17415, 17416, 17417, 17418, 17419, 17420, 17421, 17422, 17423, 17424, 17425, 17426, 17427, 17428, 17429, 17430, 17431, 17432, 17433, 17434, 17435, 17436, 17437, 17438, 17439, 17440, 17441, 17442, 17443, 17444, 17445, 17446, 17447, 17448, 17449, 17450, 17451, 17452, 17453, 17454, 17455, 17456, 17457, 17458, 17459, 17460, 17461, 17462, 17463, 17464, 17465, 17466, 17467, 17468, 17469, 17470, 17471, 17472, 17473, 17474, 17475, 17476, 17477, 17478, 17479, 17480, 17481, 17482, 17483 Extra Details: mutant; stability; hydrogen bond

Submission Details

ID: iLcnrtLf3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:49 p.m.

Version: 1

Publication Details
Prajapati RS;Lingaraju GM;Bacchawat K;Surolia A;Varadarajan R,Proteins (2003) Thermodynamic effects of replacements of Pro residues in helix interiors of maltose-binding protein. PMID:14635128
Additional Information

Study Summary

Number of data points 245
Proteins Maltose-binding periplasmic protein ; Maltose-binding periplasmic protein
Unique complexes 7
Assays/Quantities/Protocols Experimental Assay: dHcal pH:10.4 ; Experimental Assay: Tm pH:10.4 ; Experimental Assay: dHcal pH:9.9 ; Experimental Assay: Tm pH:9.9 ; Experimental Assay: dHcal pH:9.5 ; Experimental Assay: Tm pH:9.5 ; Experimental Assay: dHcal pH:9.0 ; Experimental Assay: Tm pH:9.0 ; Experimental Assay: dHcal pH:8.5 ; Experimental Assay: Tm pH:8.5 ; Experimental Assay: dHcal pH:8.0 ; Experimental Assay: Tm pH:8.0 ; Experimental Assay: dHcal pH:7.4 ; Experimental Assay: Tm pH:7.4 ; Experimental Assay: dHcal pH:7.0 ; Experimental Assay: Tm pH:7.0 ; Experimental Assay: dCp pH:6.5 ; Experimental Assay: dHcal pH:6.5 ; Experimental Assay: Tm pH:6.5 ; Experimental Assay: dCp pH:7.4, temp:63.0 C ; Experimental Assay: ddG ; Experimental Assay: Cm temp:37.0 C ; Experimental Assay: m temp:37.0 C ; Experimental Assay: dG_H2O temp:37.0 C ; Experimental Assay: Cm temp:30.0 C ; Experimental Assay: m temp:30.0 C ; Experimental Assay: dG_H2O temp:30.0 C ; Experimental Assay: Cm temp:22.0 C ; Experimental Assay: m temp:22.0 C ; Experimental Assay: dG_H2O temp:22.0 C ; Experimental Assay: Cm temp:14.0 C ; Experimental Assay: m temp:14.0 C ; Experimental Assay: dG_H2O temp:14.0 C ; Experimental Assay: Cm temp:10.0 C ; Experimental Assay: m temp:10.0 C ; Experimental Assay: dG_H2O temp:10.0 C ; Derived Quantity: dTm pH:10.4 ; Derived Quantity: dTm pH:9.9 ; Derived Quantity: dTm pH:9.5 ; Derived Quantity: dTm pH:9.0 ; Derived Quantity: dTm pH:8.5 ; Derived Quantity: dTm pH:8.0 ; Derived Quantity: dTm pH:7.4 ; Derived Quantity: dTm pH:7.0 ; Derived Quantity: dTm pH:6.5 ; Derived Quantity: ddG_H2O temp:37.0 C ; Derived Quantity: ddG_H2O temp:30.0 C ; Derived Quantity: ddG_H2O temp:22.0 C ; Derived Quantity: ddG_H2O temp:14.0 C ; Derived Quantity: ddG_H2O temp:10.0 C
Libraries Mutations for sequence KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQTRITK

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
3JYR 2009-09-22T00:00:00+0000 1.75 Crystal structures of the GacH receptor of Streptomyces glaucescens GLA.O in the unliganded form and in complex with acarbose and an acarbose homolog. Comparison with acarbose-loaded maltose binding protein of Salmonella typhimurium.
6L0Z 2019-09-27T00:00:00+0000 1.6 The crystal structure of Salmonella enterica sugar-binding protein MalE
6L3E 2019-10-10T00:00:00+0000 1.6 Crystal structure of Salmonella enterica sugar-binding protein MalE
3IO4 2009-08-13T00:00:00+0000 3.63 Huntingtin amino-terminal region with 17 Gln residues - Crystal C90
3OSQ 2010-09-09T00:00:00+0000 1.9 Maltose-bound maltose sensor engineered by insertion of circularly permuted green fluorescent protein into E. coli maltose binding protein at position 175
3OSR 2010-09-09T00:00:00+0000 2.0 Maltose-bound maltose sensor engineered by insertion of circularly permuted green fluorescent protein into E. coli maltose binding protein at position 311
3VD8 2012-01-04T00:00:00+0000 2.07 Crystal structure of human AIM2 PYD domain with MBP fusion
4FEB 2012-05-29T00:00:00+0000 2.8 Crystal Structure of Htt36Q3H-EX1-X1-C2(Beta)
4MY2 2013-09-27T00:00:00+0000 2.4 Crystal Structure of Norrin in fusion with Maltose Binding Protein
4WGI 2014-09-18T00:00:00+0000 1.85 A Single Diastereomer of a Macrolactam Core Binds Specifically to Myeloid Cell Leukemia 1 (MCL1)

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
90.7 Maltose-binding periplasmic protein P41130 MALE_PHOLU
93.9 Maltose-binding periplasmic protein P18815 MALE_KLEAE
94.3 Maltose-binding periplasmic protein P19576 MALE_SALTY
100.0 Maltose-binding periplasmic protein P0AEX9 MALE_ECOLI
100.0 Maltose-binding periplasmic protein P0AEY0 MALE_ECO57