PDBsum entry: 1eie

Hydrolase

PDB-id: 1eie

Asymmetric unit

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Protein chain

124 a.a. *

Waters ×238

* Residue conservation analysis

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Clefts Calculation

Biological unit*, dimer
(*as deduced by PQS)

PDB id:

1eie

Name:

Hydrolase

Title:

Crystal structure of f120w mutant of bovine pancreatic ribonuclease a

Structure:

Ribonuclease a. Chain: a. Engineered: yes. Mutation: yes

Source:

Bos taurus. Cattle. Organism_taxid: 9913. Organ: pancreas. Expressed in: escherichia coli. Expression_system_taxid: 562.

Biological unit:

Dimer (from PQS)

UniProt:

P61823 (RNAS1_BOVIN)

Seq:

150 a.a.

Struc:

124 a.a.*

Key:	PfamA domain
Secondary structure	CATH domain

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme class:

E.C.3.1.27.5   [IntEnz]   [ExPASy]   [KEGG]   [BRENDA]

Reaction:

Endonucleolytic cleavage to nucleoside 3'-phosphates and 3'-phosphooligonucleotides ending in C-P or U-P with 2',3'-cyclic phosphate intermediates.

Resolution:

1.40Å

R-factor:

0.218

R-free:

0.272

Authors:

E.Chatani,R.Hayashi,H.Moriyama,T.Ueki

Key ref:

E.Chatani et al. (2002). Conformational strictness required for maximum activity and stability of bovine pancreatic ribonuclease A as revealed by crystallographic study of three Phe120 mutants at 1.4 A resolution.. Protein Sci, 11, 72-81. [PubMed id: 11742124] [DOI: 10.1110/ps.ps.31102]

Date:

25-Feb-00

Release date:

13-Feb-02

Related entries:

1dp1
this is also phe120 mutant of rnase a.
1eic
this is also phe120 mutant of rnase a.
1eid
this is also phe120 mutant of rnase a.

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Key reference

DOI no: 10.1110/ps.ps.31102

Protein Sci 11:72-81 (2002)

PubMed id: 11742124

Conformational strictness required for maximum activity and stability of bovine pancreatic ribonuclease A as revealed by crystallographic study of three Phe120 mutants at 1.4 A resolution.

E.Chatani, R.Hayashi, H.Moriyama, T.Ueki.

ABSTRACT

The replacement of Phe120 with other hydrophobic residues causes a decrease in the activity and thermal stability in ribonuclease A (RNase A). To explain this, the crystal structures of wild-type RNase A and three mutants--F120A, F120G, and F120W--were analyzed up to a 1.4 A resolution. Although the overall backbone structures of all mutant samples were nearly the same as that of wild-type RNase A, except for the C-terminal region of F120G with a high B-factor, two local conformational changes were observed at His119 in the mutants. First, His119 of the wild-type and F120W RNase A adopted an A position, whereas those of F120A and F120G adopted a B position, but the static crystallographic position did not reflect either the efficiency of transphosphorylation or the hydrolysis reaction. Second, His119 imidazole rings of all mutant enzymes were deviated from that of wild-type RNase A, and those of F120W and F120G appeared to be "inside out" compared with that of wild-type RNase A. Only approximately 1 A change in the distance between N(epsilon2) of His12 and N(delta1) of His119 causes a drastic decrease in k(cat), indicating that the active site requires the strict positioning of the catalytic residues. A good correlation between the change in total accessible surface area of the pockets on the surface of the mutant enzymes and enthalpy change in their thermal denaturation also indicates that the effects caused by the replacements are not localized but extend to remote regions of the protein molecule.