Cation binding to a Bacillus (1,3-1,4)-beta-glucanase. Geometry, affinity and effect on protein stability.


Abstract

The hybrid Bacillus (1,3-1,4)-beta-glucanase H(A16-M), consisting of 16 N-terminal amino acids derived from the mature form of the B. amyloliquefaciens enzyme and of 198 C-proximal amino acids from the B. macerans enzyme, binds a calcium ion at a site at its molecular surface remote from the active center [T. Keitel, O. Simon, R. Borriss & U. Heinemann (1993) Proc. Natl Acad. Sci. USA 90, 5287-5291]. X-ray diffraction analysis at 0.22-nm resolution of crystals grown in the absence of calcium and in the presence of EDTA shows this site to be occupied by a sodium ion. Whereas the calcium ion has six oxygen atoms in its coordination sphere, two of which are from water molecules, sodium is fivefold coordinated with a fifth ligand belonging to a symmetry-related protein molecule in the crystal lattice. The affinity of H(A16-M) for calcium over sodium has been determined calorimetrically. Calcium binding stabilizes the native three-dimensional structure of the protein as shown by guanidinium chloride unfolding and thermal inactivation experiments. The enhanced enzymic activity of Bacillus beta-glucanases at elevated temperatures in the presence of calcium ions is attributed to a general stabilizing effect by the cation. Study holds ProTherm entries: 7242, 7243 Extra Details: MAC: B.macerans (1,3-1,4)-beta-glucanase calcium ion; molecular surface; active center; affinity;,thermal inactivation

Submission Details

ID: aCdYyfXX3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:33 p.m.

Version: 1

Publication Details
Keitel T;Meldgaard M;Heinemann U,Eur. J. Biochem. (1994) Cation binding to a Bacillus (1,3-1,4)-beta-glucanase. Geometry, affinity and effect on protein stability. PMID:8200344
Additional Information

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
3O5S 2010-07-28T00:00:00+0000 2.2 Crystal Structure of the endo-beta-1,3-1,4 glucanase from Bacillus subtilis (strain 168)
1AJK 1997-05-06T00:00:00+0000 1.8 CIRCULARLY PERMUTED (1-3,1-4)-BETA-D-GLUCAN 4-GLUCANOHYDROLASE CPA16M-84
1AJO 1997-05-07T00:00:00+0000 2.07 CIRCULARLY PERMUTED (1-3,1-4)-BETA-D-GLUCAN 4-GLUCANOHYDROLASE CPA16M-127
1AXK 1997-10-16T00:00:00+0000 2.1 ENGINEERED BACILLUS BIFUNCTIONAL ENZYME GLUXYN-1
1BYH 1992-12-31T00:00:00+0000 2.8 MOLECULAR AND ACTIVE-SITE STRUCTURE OF A BACILLUS (1-3,1-4)-BETA-GLUCANASE
1CPM 1994-03-11T00:00:00+0000 2.0 NATIVE-LIKE IN VIVO FOLDING OF A CIRCULARLY PERMUTED JELLYROLL PROTEIN SHOWN BY CRYSTAL STRUCTURE ANALYSIS
1CPN 1994-03-11T00:00:00+0000 1.8 NATIVE-LIKE IN VIVO FOLDING OF A CIRCULARLY PERMUTED JELLYROLL PROTEIN SHOWN BY CRYSTAL STRUCTURE ANALYSIS
1GLH 1994-11-25T00:00:00+0000 2.0 CATION BINDING TO A BACILLUS (1,3-1,4)-BETA-GLUCANASE. GEOMETRY, AFFINITY AND EFFECT ON PROTEIN STABILITY
1MAC 1994-12-22T00:00:00+0000 2.3 CRYSTAL STRUCTURE AND SITE-DIRECTED MUTAGENESIS OF BACILLUS MACERANS ENDO-1,3-1,4-BETA-GLUCANASE
1U0A 2004-07-13T00:00:00+0000 1.64 Crystal structure of the engineered beta-1,3-1,4-endoglucanase H(A16-M) in complex with beta-glucan tetrasaccharide

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
90.4 Beta-glucanase P04957 GUB_BACSU
100.0 Beta-glucanase P07980 GUB_BACAM
95.8 Beta-glucanase P23904 GUB_PAEMA