Artificial evolution of an enzyme active site: Structural studies of three highly active mutants of Escherichia coli alkaline phosphatase

Journal of Molecular Biology

Volumn 316, Issue 4, 2002, Pages 941-953

  Du, M H Le ^a   Lamoure, C ^a   Muller, B H ^a   Bulgakov, O V ^a   Lajeunesse, E ^a   Menez A ^a   Boulain, J C ^a  

^a CEA SACLAY   (France)

Author keywords

Activity enhancement; Crystallography; Mutagenesis; Phosphoseryl intermediate; Transition state

Indexed keywords

ALKALINE PHOSPHATASE; ALUMINUM FLUORIDE; BACTERIAL ENZYME; MAGNESIUM; METAL ION; MUTANT PROTEIN; PHOSPHATE; PHOSPHOSERINE; SERINE; ZINC;

ARTICLE; BACTERIAL MUTATION; CATALYSIS; CONTROLLED STUDY; COVALENT BOND; CRYSTAL STRUCTURE; ENZYME ACTIVATION; ENZYME ACTIVE SITE; ENZYME ACTIVITY; ENZYME STABILITY; ESCHERICHIA COLI; MOLECULAR EVOLUTION; NONHUMAN; PRIORITY JOURNAL; WILD TYPE;

BACTERIA (MICROORGANISMS); ESCHERICHIA COLI; NEGIBACTERIA; TRIXIS;

EID: 0036290280     PISSN: 00222836     EISSN: None     Source Type: Journal    
DOI: 10.1006/jmbi.2001.5384     Document Type: Article

References (44)

1. Structure and mechanism of alkaline phosphatase
2. Partial sequencing of human adult, human fetal, and bovine intestinal alkaline phosphatases: Comparison with the human placental and liver isozymes
3. Improving Escherichia coli alkaline phosphatase efficacy by additional mutations inside and outside the catalytic pocket
4. Phosphate incorporation into alkaline phosphatase of Escherichia coli
5. Structure of alkaline phosphatases
6. Reaction mechanism of alkaline phosphatase based on crystal structures. Two-metal ion catalysis
7. 3-D structure of a mutant (Asp101 → Ser) of E. coli alkaline phosphatase with higher catalytic activity
8. Mutagenesis of conserved residues within the active site of Escherichia coli alkaline phosphatase yields enzymes with increased kcat
9. Enhanced catalysis by active-site mutagenesis at aspartic acid 153 in Escherichia coli alkaline phosphatase
10. A water-mediated salt link in the catalytic site of Escherichia coli alkaline phosphatase may influence activity
11. Magnesium in the active site of Escherichia coli alkaline phosphatase is important for both structural stabilization and catalysis
12. Why are mammalian alkaline phosphatases much more active than bacterial alkaline phosphatases?
13. The mechanism of the alkaline phosphatase reaction: Insights from NMR, crystallography and site-specific mutagenesis
14. A revised mechanism for the alkaline phosphatase reaction involving three metal ions
15. 31P nuclear magnetic resonance of phosphoenzyme intermediates of alkaline phosphatase
16. 31P nuclear magnetic resonance study of alkaline phosphatase: The role of inorganic phosphate in limiting the enzyme turnover rate at alkaline pH
17. 113Cd NMR
18. Trapping and visualization of a covalent enzyme-phosphate intermediate
19. Crystallographic analysis of reversible metal binding observed in a mutant (Asp153 → Gly) of Escherichia coli alkaline phosphatase
20. 3-D structure of the D153G mutant of Escherichia coli alkaline phosphatase: An enzyme with weaker magnesium binding and increased catalytic activity
21. Crystal structure of alkaline phosphatase from human placenta at 1.8 Å resolution. Implication for a substrate specificity
22. Structural evidences for a functional role of human tissue non-specific alkaline phosphatase in bone mineralization
23. Mutations at positions 153 and 328 in Escherichia coli alkaline phosphatase provide insight towards the structure and function of mammalian and yeast alkaline phosphatases
24. Conversion of a magnesium binding site into a zinc binding site by a single amino acid substitution in Escherichia coli alkaline phosphatase
25. Kinetic and X-ray structural studies of three mutant E. coli alkaline phosphatases: Insights into the catalytic mechanism without the nucleophile Ser102
26. Kinetics and crystal structure of a mutant Escherichia coli alkaline phosphatase (Asp-369 → Asn): A mechanism involving one zinc per active site
27. Cloning and expression of the bovine intestinal alkaline phosphatase gene: Biochemical characterization of the recombinant enzyme
28. AMoRe: An automated package for molecular replacement
29. A model of the transition state in the alkaline phosphatase reaction
30. Recombinant single-chain Fv antibody fragment-alkaline phosphatase conjugate for one-step immunodetection in molecular hybridization
31. Insertion of a disulfide-containing neurotoxin into E. coli alkaline phosphatase: The hybrid retains both biological activities
32. The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts
33. Cleavage of structural proteins during the assembly of the head of the bacteriophage T4
34. Interaction of alkaline phosphatase of E. coli with metal ions and chelating agents
35. A fine structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. I. Purification and characterization of alkaline phosphatase
36. AlF3 mimics the transition state of protein phosphorylation in the crystal structure of nucleoside diphosphate kinase and MgADP
37. Structure at 1.65 Å of RhoA and its GTPase-activationg protein in complex with a transition-state analogue
38. Automatic processing of rotation diffraction data from crystals of initially unknown symmetry and cell constants
39. Processing of X-ray diffraction data collected in oscillation mode
40. TheCCP4 suite: Programs for proteins crystallography
41. PROCHECK: A program to check the stereochemical quality of protein structures
42. Phosphocholine binding immunoglobuline Fab McPC603 an X-ray study at 2.7 Å