PDBsum entry 4xoj

Hydrolase

PDB id: 4xoj

Contents

Protein chains

223 a.a.

13 a.a.

Ligands

SO4

EDO ×7

NH4

Metals

_CA

_NA ×3

Waters ×397

PDB id:

4xoj

Name:

Hydrolase

Title:

Structure of bovine trypsin in complex with analogues of sunflower inhibitor 1 (sfti-1)

Structure:

Cationic trypsin. Chain: a. Synonym: beta-trypsin. Trypsin inhibitor 1. Chain: b. Fragment: unp residues 40-52. Synonym: sfti-1. Engineered: yes

Source:

Bos taurus. Bovine. Organism_taxid: 9913. Synthetic: yes. Helianthus annuus. Common sunflower. Organism_taxid: 4232

Resolution:

0.91Å R-factor: 0.104 R-free: 0.114

Authors:

P.Golik,S.Malicki,P.Grudnik,N.Karna,D.Debowski,A.Legowska,B.Wladyka, A.Gitlin,K.Brzozowski,G.Dubin,K.Rolka

Key ref:

N.Karna et al. (2015). Investigation of Serine-Proteinase-Catalyzed Peptide Splicing in Analogues of Sunflower Trypsin Inhibitor 1 (SFTI-1). Chembiochem, 16, 2036-2045. PubMed id: 26212347 DOI: 10.1002/cbic.201500296

Date:

16-Jan-15 Release date: 12-Aug-15

Protein chain

P00760  (TRY1_BOVIN) -  Serine protease 1 from Bos taurus

Seq: 246 a.a.

Struc: 223 a.a.

Protein chain

Q4GWU5  (SFTI1_HELAN) -  Trypsin inhibitor 1 from Helianthus annuus

Seq: 56 a.a.

Struc: 13 a.a.

Enzyme reactions

• Enzyme class: Chain A: E.C.3.4.21.4 - trypsin.
• Reaction: Preferential cleavage: Arg-|-Xaa, Lys-|-Xaa.

DOI no: 10.1002/cbic.201500296

Chembiochem 16:2036-2045 (2015)

PubMed id: 26212347

Investigation of Serine-Proteinase-Catalyzed Peptide Splicing in Analogues of Sunflower Trypsin Inhibitor 1 (SFTI-1).

N.Karna, A.Łęgowska, S.Malicki, D.Dębowski, P.Golik, A.Gitlin, P.Grudnik, B.Wladyka, K.Brzozowski, G.Dubin, K.Rolka.

ABSTRACT

Serine-proteinase-catalyzed peptide splicing was demonstrated in analogues of the trypsin inhibitor SFTI-1: both single peptides and two-peptide chains (C- and N-terminal peptide chains linked by a disulfide bridge). In the second series, peptide splicing with catalytic amount of proteinase was observed only when formation of acyl-enzyme intermediate was preceded by hydrolysis of the substrate Lys-Ser peptide bond. Here we demonstrate that with an equimolar amount of the proteinase, splicing occurs in all the two-peptide-chain analogues. This conclusion was supported by high resolution crystal structures of selected analogues in complex with trypsin. We showed that the process followed a direct transpeptidation mechanism. Thus, the acyl-enzyme intermediate was formed and was immediately used for a new peptide bond formation; products associated with the hydrolysis of the acyl-enzyme were not observed. The peptide splicing was sequence- not structure-specific.