PDBsum entry 4d6c

Hydrolase

PDB id: 4d6c

Contents

Protein chain

586 a.a.

Ligands

EDO ×2

Waters ×818

PDB id:

4d6c

Name:

Hydrolase

Title:

Crystal structure of a family 98 glycoside hydrolase catalytic module (sp3gh98)(l19 mutant)

Structure:

Glycoside hydrolase. Chain: a. Fragment: catalytic domain, unp residues 423-1005. Engineered: yes. Mutation: yes

Source:

Streptococcus pneumoniae. Organism_taxid: 406556. Strain: sp3-bs71. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

1.59Å R-factor: 0.156 R-free: 0.179

Authors:

D.H.Kwan,I.Constantinescu,R.Chapanian,M.A.Higgins,E.Samain, A.B.Boraston,J.N.Kizhakkedathu,S.G.Withers

Key ref:

D.H.Kwan et al. (2015). Toward Efficient Enzymes for the Generation of Universal Blood through Structure-Guided Directed Evolution. J Am Chem Soc, 137, 5695-5705. PubMed id: 25870881 DOI: 10.1021/ja5116088

Date:

11-Nov-14 Release date: 26-Nov-14

Protein chain

A5LBQ0  (A5LBQ0_STREE) -

Enzyme reactions

• Enzyme class: E.C.3.2.1.102 - blood-group-substance endo-1,4-beta-galactosidase.
• Reaction: Endohydrolysis of 1,4-beta-D-galactosidic linkages in blood group A and B substances.

DOI no: 10.1021/ja5116088

J Am Chem Soc 137:5695-5705 (2015)

PubMed id: 25870881

Toward Efficient Enzymes for the Generation of Universal Blood through Structure-Guided Directed Evolution.

D.H.Kwan, I.Constantinescu, R.Chapanian, M.A.Higgins, M.P.Kötzler, E.Samain, A.B.Boraston, J.N.Kizhakkedathu, S.G.Withers.

ABSTRACT

Blood transfusions are critically important in many medical procedures, but the presence of antigens on red blood cells (RBCs, erythrocytes) means that careful blood-typing must be carried out prior to transfusion to avoid adverse and sometimes fatal reactions following transfusion. Enzymatic removal of the terminal N-acetylgalactosamine or galactose of A- or B-antigens, respectively, yields universal O-type blood, but is inefficient. Starting with the family 98 glycoside hydrolase from Streptococcus pneumoniae SP3-BS71 (Sp3GH98), which cleaves the entire terminal trisaccharide antigenic determinants of both A- and B-antigens from some of the linkages on RBC surface glycans, through several rounds of evolution, we developed variants with vastly improved activity toward some of the linkages that are resistant to cleavage by the wild-type enzyme. The resulting enzyme effects more complete removal of blood group antigens from cell surfaces, demonstrating the potential for engineering enzymes to generate antigen-null blood from donors of various types.