PDBsum entry 3i78

Hydrolase

PDB id

3i78

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Contents

			Protein chain

					229 a.a. *

			Ligands

					BEN


					SO4 ×4

			Metals

					_NA

* Residue conservation analysis

PDB id:

3i78

Links

Name:

Hydrolase

Title:

35/99/170/186/220-loops of fxa in sgt

Structure:

Trypsin. Chain: a. Synonym: sgt. Engineered: yes. Mutation: yes. Other_details: 35/99/170/186/220-loops of fxa in sgt

Source:

Streptomyces griseus. Organism_taxid: 1911. Gene: sprt. Expressed in: bacillus subtilis. Expression_system_taxid: 1423.

Resolution:

3.00Å

R-factor:

0.211

R-free:

0.275

Authors:

M.J.Page,E.Di Cera

Key ref:

M.J.Page and E.Di Cera (2010). Combinatorial enzyme design probes allostery and cooperativity in the trypsin fold. J Mol Biol, 399, 306-319. PubMed id: 20399789

Date:

08-Jul-09

Release date:

02-Jun-10

PROCHECK

	Headers

	References

Protein chain

P00775 (TRYP_STRGR) - Trypsin from Streptomyces griseus

Seq: Struc:					259 a.a. 229 a.a.*

Key:

PfamA domain

Secondary structure

CATH domain

* PDB and UniProt seqs differ at 27 residue positions (black crosses)



		Enzyme reactions

Enzyme class:

E.C.3.4.21.4 - trypsin.

[IntEnz] [ExPASy] [KEGG] [BRENDA]

Reaction:

Preferential cleavage: Arg-|-Xaa, Lys-|-Xaa.

	J Mol Biol 399:306-319 (2010)
PubMed id: 20399789

Combinatorial enzyme design probes allostery and cooperativity in the trypsin fold.
M.J.Page, E.Di Cera.

ABSTRACT

Converting one enzyme into another is challenging due to the uneven distribution of important amino acids for function in both protein sequence and structure. We report a strategy for protein engineering allowing an organized mixing and matching of genetic material that leverages lower throughput with increased quality of screens. Our approach successfully tested the contribution of each surface-exposed loop in the trypsin fold alone and the cooperativity of their combinations towards building the substrate selectivity and Na(+)-dependent allosteric activation of the protease domain of human coagulation factor Xa into a bacterial trypsin. As the created proteases lack additional protein domains and protein co-factor activation mechanism requisite for the complexity of blood coagulation, they are stepping-stones towards further understanding and engineering of artificial clotting factors.

Literature references that cite this PDB file's key reference

PubMed id

Reference

21406063

C.S.Craik, M.J.Page, and E.L.Madison (2011).
Proteases as therapeutics.

Biochem J, 435, 1.

21368156

S.Rana, N.Pozzi, L.A.Pelc, and E.Di Cera (2011).
Redesigning allosteric activation in an enzyme.

Proc Natl Acad Sci U S A, 108, 5221-5225.

The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time.