Unfolding studies on soybean agglutinin and concanavalin a tetramers: a comparative account.


Abstract

The unfolding pathway of two very similar tetrameric legume lectins soybean agglutinin (SBA) and Concanavalin A (ConA) were determined using GdnCl-induced denaturation. Both proteins displayed a reversible two-state unfolding mechanism. The analysis of isothermal denaturation data provided values for conformational stability of the two proteins. It was found that the DeltaG of unfolding of SBA was much higher than ConA at all the temperatures at which the experiments were done. ConA had a T(g) 18 degrees C less than SBA. The higher conformational stability of SBA in comparison to ConA is largely due to substantial differences in their degrees of subunit interactions. Ionic interactions at the interface of the two proteins especially at the noncanonical interface seem to play a significant role in the observed stability differences between these two proteins. Furthermore, SBA is a glycoprotein with a GlcNac2Man9 chain attached to Asn-75 of each subunit. The sugar chain in SBA lies at the noncanonical interface of the protein, and it is found to interact with the amino acid residues in the adjacent noncanonical interface. These interactions further stabilize SBA with respect to ConA, which is not glycosylated. Study holds ProTherm entries: 19171, 19172, 19173, 19174, 19175, 19176, 19177, 19178, 19179, 19180, 19181, 19182, 19183, 19184, 19185, 19186, 19187, 19188, 19189, 19190, 19191, 19192, 19193 Extra Details: lectins; two-state unfolding; noncanonical interface

Submission Details

ID: hpi5vyYB3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:51 p.m.

Version: 1

Publication Details
Sinha S;Mitra N;Kumar G;Bajaj K;Surolia A,Biophys. J. (2005) Unfolding studies on soybean agglutinin and concanavalin a tetramers: a comparative account. PMID:15542553
Additional Information

Study Summary

Number of data points 48
Proteins Lectin ; Lectin ; Concanavalin-A ; Concanavalin-A
Unique complexes 2
Assays/Quantities/Protocols Experimental Assay: dCp ; Experimental Assay: Tm ; Experimental Assay: dHvH ; Experimental Assay: m temp:312 K ; Experimental Assay: dG_H2O temp:312 K ; Experimental Assay: m temp:294 K ; Experimental Assay: dG_H2O temp:294 K ; Experimental Assay: m temp:291 K ; Experimental Assay: dG_H2O temp:291 K ; Experimental Assay: m temp:285 K ; Experimental Assay: dG_H2O temp:285 K ; Experimental Assay: m temp:280 K ; Experimental Assay: dG_H2O temp:280 K ; Experimental Assay: m temp:323 K ; Experimental Assay: dG_H2O temp:323 K ; Experimental Assay: m temp:318 K ; Experimental Assay: dG_H2O temp:318 K ; Experimental Assay: m temp:313 K ; Experimental Assay: dG_H2O temp:313 K ; Experimental Assay: m temp:306 K ; Experimental Assay: dG_H2O temp:306 K ; Experimental Assay: m temp:303 K ; Experimental Assay: dG_H2O temp:303 K ; Experimental Assay: m temp:300 K ; Experimental Assay: dG_H2O temp:300 K ; Experimental Assay: m temp:298 K ; Experimental Assay: dG_H2O temp:298 K ; Experimental Assay: m temp:295 K ; Experimental Assay: dG_H2O temp:295 K ; Experimental Assay: m temp:293 K ; Experimental Assay: dG_H2O temp:293 K ; Experimental Assay: m temp:288 K ; Experimental Assay: dG_H2O temp:288 K ; Experimental Assay: m temp:283 K ; Experimental Assay: dG_H2O temp:283 K ; Experimental Assay: m temp:281 K ; Experimental Assay: dG_H2O temp:281 K
Libraries Mutations for sequence ADTIVAVELDTYPNTDIGDPSYPHIGIDIKSVRSKKTAKWNMQNGKVGTAHIIYNSVDKRLSAVVSYPNADSATVSYDVDLDNVLPEWVRVGLSASTGLYKETNTILSWSFTSKLKSNSTHETNALHFMFNQFSKDQKDLILQGDATTGTDGNLELTRVSSNGSPQGSSVGRALFYAPVHIWESSAVVASFEATFTFLIKSPDSHPADGIAFFISNIDSSIPSGSTGRLLGLFPDAN ; Mutations for sequence AETVSFSWNKFVPKQPNMILQGDAIVTSSGKLQLNKVDENGTPKPSSLGRALYSTPIHIWDKETGSVASFAASFNFTFYAPDTKRLADGLAFFLAPIDTKPQTHAGYLGLFNENESGDQVVAVEFDTFRNSWDPPNPHIGINVNSIRSIKTTSWDLANNKVAKVLITYDASTSLLVASLVYPSQRTSNILSDVVDLKTSLPEWVRIGFSAATGLDIPGESHDVLSWSFASNLPHASSNIDPLDLTSFVLHEAI

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
5WEY 2017-09-13 Joint X-ray/neutron structure of Concanavalin A with alpha1-2 D-mannobiose
2UU8 2007-07-31 0.94 X-ray structure of Ni, Ca concanavalin A at Ultra-high resolution (0. 94A)
1NLS 1997-11-26 0.94 CONCANAVALIN A AND ITS BOUND SOLVENT AT 0.94A RESOLUTION
1JBC 1997-02-12 1.15 CONCANAVALIN A
1I3H 2001-07-25 1.2 CONCANAVALIN A-DIMANNOSE STRUCTURE
5O6N 2017-12-13 1.35 High pressure flash cooled concanavalin A
5Z5P 2018-11-07 1.4 Crystal structure of ConA-R3M
2OVU 2007-10-23 1.5 Crystal structure of a lectin from Canavalia gladiata (CGL) in complex with man1-2man-OMe
1GKB 2001-08-20 1.56 CONCANAVALIN A, NEW CRYSTAL FORM
1SCS 1994-05-31 1.6 HIGH-RESOLUTION STRUCTURES OF SINGLE-METAL-SUBSTITUTED CONCANAVALIN A: THE CO,CA-PROTEIN AT 1.6 ANGSTROMS AND THE NI,CA-PROTEIN AT 2.0 ANGSTROMS
5YGM 2017-10-25 1.6 Monomeric structure of concanavalin A at pH 7.5 from Carnivalia ensiformis
4K21 2014-04-09 1.6 Crystal structure of Canavalia boliviana lectin in complex with Xman
3QLQ 2012-02-08 1.7 Crystal structure of Concanavalin A bound to an octa-alpha-mannosyl-octasilsesquioxane cluster
1DQ0 2000-01-19 1.7 Locked, metal-free concanavalin A, a minor species in solution
4CZS 2015-04-22 1.73 Discovery of Glycomimetic Ligands via Genetically-encoded Library of Phage displaying Mannose-peptides
2A7A 2005-07-19 1.75 On the Routine Use of Soft X-Rays in Macromolecular Crystallography, Part III- The Optimal Data Collection Wavelength
2CYF 2006-06-13 1.8 The Crystal Structure of Canavalia Maritima Lectin (ConM) in Complex with Trehalose and Maltose
3D4K 2009-03-03 1.8 Concanavalin A Complexed to a Synthetic Analog of the Trimannoside
1QNY 2000-04-30 1.8 X-ray refinement of D2O soaked crystal of concanavalin A
1ENR 1996-08-17 1.83 CO-CRYSTALS OF DEMETALLIZED CONCANAVALIN A WITH ZINC AND CALCIUM HAVING A ZINC ION BOUND IN THE S1 SITE AND A CALCIUM ION BOUND IN THE S2 SITE
5Z5Y 2018-05-30 1.89 Crystal structure of ConA-R4M
4I30 2012-12-12 1.89 Crystal structure of Canavalia maritima seeds lectin (ConM) co-crystalized with gamma-aminobutyric acid (GABA) and soaked with adenine
4P9Y 2014-08-27 1.89 Structure of ConA/Rh4man
1JN2 2003-07-01 1.9 Crystal Structure of meso-tetrasulphonatophenyl porphyrin complexed with Concanavalin A
1NXD 2004-03-30 1.9 Crystal structure of MnMn Concanavalin A
1DQ6 2000-01-19 1.9 Manganese;Manganese concanavalin A at pH 7.0
1JW6 2001-09-26 1.93 Crystal Structure of the Complex of Concanavalin A and Hexapeptide
6H2M 2019-03-13 1.93 Long wavelength Mesh&Collect native SAD phasing on microcrystals
2CTV 1993-10-31 1.95 HIGH RESOLUTION CRYSTALLOGRAPHIC STUDIES OF NATIVE CONCANAVALIN A USING RAPID LAUE DATA COLLECTION METHODS AND THE INTRODUCTION OF A MONOCHROMATIC LARGE-ANGLE OSCILLATION TECHNIQUE (LOT)
2P2K 2007-10-23 1.98 Crystal structure of a lectin from Canavalia gladiata seeds (CGL) in complex with man1-4man-OMe
1DQ5 2000-01-19 2.0 Manganese;Manganese concanavalin A at pH 5.0
2CY6 2006-06-13 2.0 Crystal structure of ConM in complex with trehalose and maltose
1CON 1994-01-31 2.0 THE REFINED STRUCTURE OF CADMIUM SUBSTITUTED CONCANAVALIN A AT 2.0 ANGSTROMS RESOLUTION
4P14 2015-04-15 2.0 Crystal structure of Canavalia brasiliensis (ConBr) complexed with adenine
1SCR 1994-05-31 2.0 HIGH-RESOLUTION STRUCTURES OF SINGLE-METAL-SUBSTITUTED CONCANAVALIN A: THE CO,CA-PROTEIN AT 1.6 ANGSTROMS AND THE NI,CA-PROTEIN AT 2.0 ANGSTROMS
2CNA 1977-03-16 2.0 THE COVALENT AND THREE-DIMENSIONAL STRUCTURE OF CONCANAVALIN A, IV.ATOMIC COORDINATES,HYDROGEN BONDING,AND QUATERNARY STRUCTURE
1GIC 1997-08-20 2.0 CONCANAVALIN A COMPLEXED WITH METHYL ALPHA-D-GLUCOPYRANOSIDE
5CNA 1994-05-31 2.0 REFINED STRUCTURE OF CONCANAVALIN A COMPLEXED WITH ALPHA-METHYL-D-MANNOPYRANOSIDE AT 2.0 ANGSTROMS RESOLUTION AND COMPARISON WITH THE SACCHARIDE-FREE STRUCTURE
1QDC 1999-10-14 2.0 MAN(APLHA1-6)MAN(ALPHA1-O)METHYL CONCANAVALIN A COMPLEX
4PF5 2014-12-17 2.04 Crystal structure of Concanavalin A complexed with a synthetic derivative of high-mannose chain
5Z5N 2018-11-07 2.04 Crystal structure of ConA-R1M
1DQ2 2000-01-19 2.05 Unlocked metal-free concanavalin A
4P9X 2014-08-27 2.06 Structure of ConA/Rh3Glu complex
3NWK 2011-07-13 2.09 A second C2221 form of concanavalin A (Canavalia ensiformis)
6GW9 2018-09-05 2.1 Concanavalin A structure determined with data from the EuXFEL, the first MHz free electron laser
3JU9 2010-12-08 2.1 Crystal structure of a lectin from Canavalia brasiliensis seed (ConBr) complexed with alpha-aminobutyric acid
2EF6 2007-10-30 2.1 Canavalia gladiata lectin complexed with Man1-3Man-OMe
4P9W 2014-08-27 2.11 Structure of ConA/Rh3Man
4PCR 2015-04-22 2.15 Crystal structure of Canavalia brasiliensis seed lectin (ConBr) complexed with Gamma-Aminobutyric Acid (GABA)
4H55 2012-10-31 2.15 Crystal structure of Canavalia brasiliensis seed lectin (ConBr) in complex with beta-d-ribofuranose
1DQ1 2000-01-19 2.15 Calcium;Calcium concanavalin A
3SNM 2012-08-01 2.15 Crystal structure of a lectin from Canavalia maritima seeds complexed with Indole-3-Acetic Acid
2YZ4 2008-02-05 2.2 The neutron structure of concanavalin A at 2.2 Angstroms
5U3E 2017-08-23 2.3 Crystal Structure of Native Lectin from Canavalia bonariensis Seeds (CaBo) complexed with alpha-methyl-D-mannoside
4L8Q 2014-05-21 2.3 Crystal structure of Canavalia grandiflora seed lectin complexed with X-Man.
4K1Z 2014-04-09 2.3 Crystal structure of Canavalia boliviana lectin in complex with Man1-4Man-OMe
1CVN 1996-10-14 2.3 CONCANAVALIN A COMPLEXED TO TRIMANNOSIDE
5Z5L 2019-04-10 2.3 Crystal structure of ConA-R5M
1WUV 2006-04-18 2.3 Crystal structure of native Canavalia gladiata lectin (CGL): a tetrameric ConA-like lectin
2D7F 2006-12-19 2.31 Crystal structure of A lectin from canavalia gladiata seeds complexed with alpha-methyl-mannoside and alpha-aminobutyric acid
1ONA 1997-09-17 2.35 CO-CRYSTALS OF CONCANAVALIN A WITH METHYL-3,6-DI-O-(ALPHA-D-MANNOPYRANOSYL)-ALPHA-D-MANNOPYRANOSIDE
2ENR 1999-02-16 2.35 CO-CRYSTALS OF DEMETALLIZED CONCANAVALIN A WITH CADMIUM HAVING A CADMIUM ION BOUND IN BOTH THE S1 SITE AND THE S2 SITE
2G4I 2007-02-20 2.4 Anomalous substructure of Concanavalin A
1C57 2000-05-08 2.4 DIRECT DETERMINATION OF THE POSITIONS OF DEUTERIUM ATOMS OF BOUND WATER IN CONCANAVALIN A BY NEUTRON LAUE CRYSTALLOGRAPHY
3ENR 2000-09-18 2.4 ZINC-CALCIUM CONCANAVALIN A AT PH 6.15
1HQW 2003-10-07 2.4 CRYSTAL STRUCTURE OF THE COMPLEX OF CONCANAVALIN A WITH A TRIPEPTIDE YPY
3CNA 1976-10-11 2.4 STRUCTURE OF CONCANAVALIN A AT 2.4 ANGSTROMS RESOLUTION
1SBF 1998-04-22 2.43 SOYBEAN AGGLUTININ
4K1Y 2014-04-09 2.5 Crystal structure of Canavalia boliviana lectin in complex with Man1-3Man-OMe
1XQN 2004-11-02 2.5 The 15k neutron structure of saccharide-free concanavalin A
1ENQ 1996-08-17 2.5 CO-CRYSTALS OF DEMETALLIZED CONCANAVALIN A WITH ZINC HAVING A ZINC ION BOUND IN THE S1 SITE
1APN 1996-04-03 2.5 THE CRYSTALLOGRAPHIC STRUCTURE OF METAL-FREE CONCANAVALIN A AT 2.5 ANGSTROMS RESOLUTION
1G9F 2001-06-13 2.5 CRYSTAL STRUCTURE OF THE SOYBEAN AGGLUTININ IN A COMPLEX WITH A BIANTENNARY BLOOD GROUP ANTIGEN ANALOG
1SBD 1998-04-22 2.52 SOYBEAN AGGLUTININ COMPLEXED WITH 2,4-PENTASACCHARIDE
5F5Q 2016-10-26 2.52 Crystal structure of Canavalia virosa lectin in complex with alpha-methyl-mannoside
4DPN 2013-03-06 2.55 Crystal Structure of ConM Complexed with Resveratrol
5ZAC 2018-05-30 2.59 Crystal structure of ConA-R2M
2SBA 1998-12-09 2.6 SOYBEAN AGGLUTININ COMPLEXED WITH 2,6-PENTASACCHARIDE
5BYN 2015-11-18 2.65 Canavalia maritima lectin complexed with synthetic selenoamino acid
1QGL 1999-05-20 2.66 Room temperature structure of concanavalin A complexed to bivalent ligand
1TEI 1997-08-20 2.7 STRUCTURE OF CONCANAVALIN A COMPLEXED TO BETA-D-GLCNAC (1,2)ALPHA-D-MAN-(1,6)[BETA-D-GLCNAC(1,2)ALPHA-D-MAN (1,6)]ALPHA-D-MAN
4D69 2014-11-19 2.7 SOYBEAN AGGLUTININ FROM GLYCINE MAX IN COMPLEX WITH THE ANTIGEN Tn
1CES 1997-02-15 2.7 CRYSTALS OF DEMETALLIZED CONCANAVALIN A SOAKED WITH ZINC HAVE A ZINC ION BOUND IN THE S1 SITE
1VAM 1997-01-11 2.75 CONCANAVALIN A COMPLEX WITH 4'-NITROPHENYL-ALPHA-D-MANNOPYRANOSIDE
1JYC 2002-09-12 2.75 CONCANAVALIN A/15-mer PEPTIDE COMPLEX
1JUI 2002-08-24 2.75 CONCANAVALIN A-CARBOHYDRATE MIMICKING 10-MER PEPTIDE COMPLEX
1JYI 2002-09-12 2.75 CONCANAVALIN A/12-MER PEPTIDE COMPLEX
1BXH 1998-10-07 2.75 CONCANAVALIN A COMPLEXED TO METHYL ALPHA1-2 MANNOBIOSIDE
1CJP 1997-10-15 2.78 CONCANAVALIN A COMPLEX WITH 4'-METHYLUMBELLIFERYL-ALPHA-D-GLUCOPYRANOSIDE
1ENS 1996-08-17 2.8 CRYSTALS OF DEMETALLIZED CONCANAVALIN A SOAKED WITH COBALT HAVING A COBALT ION BOUND IN THE S1 SITE
1QDO 1999-10-14 2.8 MAN(APLHA1-3)MAN(ALPHA1-O)METHYL CONCANAVALIN A COMPLEX
1SBE 1998-04-22 2.8 SOYBEAN AGGLUTININ FROM GLYCINE MAX
6AHG 2018-11-21 2.83 Trimeric structure of concanavalin A from Canavalia ensiformis
1DQ4 2000-01-19 2.9 A transient unlocked concanavalin A structure with MN2+ bound in the transition metal ion binding site S1 and an empty calcium binding site S2
1AZD 1998-04-29 3.0 CONCANAVALIN FROM CANAVALIA BRASILIENSIS
1JOJ 2001-12-19 3.0 CONCANAVALIN A-HEXAPEPTIDE COMPLEX
1VAL 1997-01-11 3.0 CONCANAVALIN A COMPLEX WITH 4'-NITROPHENYL-ALPHA-D-GLUCOPYRANOSIDE
4TZD 2015-07-22 3.2 Crystal structure of Canavalia maritima lectin (ConM) complexed with interleukin - 1 beta primer
1CN1 1982-03-04 3.2 CRYSTAL STRUCTURE OF DEMETALLIZED CONCANAVALIN A. THE METAL-BINDING REGION
4TYS 2015-07-22 3.25 Crystal structure of Canavalia maritima lectin (ConM) complexed with a dinucleotide
4K20 2014-04-09 3.4 Crystal structure of Canavalia boliviana lectin

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Lectin P05046 LEC_SOYBN
91.5 Concanavalin-A P58906 LECA_CANBN
98.6 Concanavalin-A P14894 CONA_CANGL
100.0 Concanavalin-A P02866 CONA_CANEN
94.1 Concanavalin-A A0A067XG71 LECA_CANGR
97.5 Concanavalin-A A0A023GPI8 LECA_CANBL
98.3 Concanavalin-A P81460 CONA_CANLI
98.3 Concanavalin-A P55915 CONA_CANBR
98.3 Concanavalin-A C0HJY1 CONV_CANCT
98.3 Concanavalin-A P81461 CONA_CANCT