Effect of linker segments on the stability of epithelial cadherin Domain 2.


Abstract

Epithelial cadherin is a transmembrane protein that is essential in calcium-dependent cell-cell recognition and adhesion. It contains five independently folded globular domains in its extracellular region. Each domain has a seven-strand beta-sheet immunoglobulin fold. Short seven-residue peptide segments connect the globular domains and provide oxygens to chelate calcium ions at the interface between the domains (Nagar et al., Nature 1995;380:360-364). Recently, stability studies of ECAD2 (Prasad et al., Biochemistry 2004;43:8055-8066) were undertaken with the motivation that Domain 2 is a representative domain for this family of proteins. The definition of a domain boundary is somewhat arbitrary; hence, it was important to examine the effect of the adjoining linker regions that connect Domain 2 to the adjacent domains. Present studies employ temperature-denaturation and proteolytic susceptibility to provide insight into the impact of these linkers on Domain 2. The significant findings of our present study are threefold. First, the linker segments destabilize the core domain in the absence of calcium. Second, the destabilization due to addition of the linker segments can be partially reversed by the addition of calcium. Third, sodium chloride stabilizes all constructs. This result implies that electrostatic repulsion is a contributor to destabilization of the core domain by addition of the linkers. Thus, the context of Domain 2 within the whole molecule affects its thermodynamic characteristics. Study holds ProTherm entries: 22049, 22050, 22051, 22052, 22053, 22054, 22055, 22056, 22057, 22058, 22059, 22060, 22061, 22062, 22063, 22064 Extra Details: domain 2 (residues 107-212) thermodynamics; calcium binding; modular domains; circular dichroism; thermal denaturation

Submission Details

ID: qcysxasn3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:53 p.m.

Version: 1

Publication Details
Prasad A;Zhao H;Rutherford JM;Housley N;Nichols C;Pedigo S,Proteins (2006) Effect of linker segments on the stability of epithelial cadherin Domain 2. PMID:16287100
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
1OP4 2004-03-16 Solution Structure of Neural Cadherin Prodomain
1NCG 1995-07-10 2.1 STRUCTURAL BASIS OF CELL-CELL ADHESION BY CADHERINS
1NCI 1995-07-10 2.1 STRUCTURAL BASIS OF CELL-CELL ADHESION BY CADHERINS
1NCH 1995-07-10 2.1 STRUCTURAL BASIS OF CELL-CELL ADHESION BY CADHERINS
4NUQ 2014-09-24 2.12 Crystal structure of mouse N-cadherin EC1-2 W2F
4NUP 2014-09-24 2.7 Crystal structure of mouse N-cadherin EC1-2 with AA insertion between residues 2 and 3
2QVI 2008-08-12 3.01 Crystal structure of N-cadherin domains EC12
3Q2W 2011-02-23 3.2 Crystal structure of mouse N-cadherin ectodomain
4NUM 2014-09-24 3.3 Crystal structure of mouse N-cadherin EC1-2 A78SI92M
1NCJ 1999-03-18 3.4 N-CADHERIN, TWO-DOMAIN FRAGMENT

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
98.3 Cadherin-2 O55075 CADH2_CRIGR
99.1 Cadherin-2 P19534 CADH2_BOVIN
99.1 Cadherin-2 B2KI42 CADH2_RHIFE
99.1 Cadherin-2 B4USZ0 CADH2_OTOGA
98.1 Cadherin-2 Q5R9X1 CADH2_PONAB
98.1 Cadherin-2 P19022 CADH2_HUMAN
98.1 Cadherin-2 B0KW95 CADH2_CALJA
99.1 Cadherin-2 Q9Z1Y3 CADH2_RAT
100.0 Cadherin-2 P15116 CADH2_MOUSE
90.7 Cadherin-2 P20310 CAD2B_XENLA
95.4 Cadherin-2 P10288 CADH2_CHICK