PMID- 8399158
OWN - NLM
STAT- MEDLINE
DCOM- 19931102
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 32
IP  - 38
DP  - 1993 Sep 28
TI  - Engineering a cysteine ligand into the zinc binding site of human carbonic 
      anhydrase II.
PG  - 9896-900
AB  - Substitution of cysteine for threonine-199, the amino acid which hydrogen bonds 
      with zinc-bound hydroxide in wild-type carbonic anhydrase II (CAII), leads to the 
      formation of a new His3Cys zinc coordination polyhedron. The optical absorption 
      spectrum of the Co(2+)-substituted threonine-199-->cysteine (T199C) variant and 
      the three-dimensional structure [Ippolito, J. A., & Christianson, D. W. (1993) 
      Biochemistry (following paper in this issue)] indicate that the new thiolate side 
      chain coordinates to the metal ion, displacing the metal-bound solvent molecule. 
      The engineered thiolate ligand increases zinc binding (4-fold) and decreases 
      catalytic activity substantially (approximately 10(3)-fold) but not completely. 
      However, this residual activity is due to an active species containing a 
      zinc-bound solvent ligand with the cysteine-199 side chain occupying an alternate 
      conformation. The equilibrium between these conformers reflects the energetic 
      balance between the formation of the zinc-thiolate bond and structural 
      rearrangements in the Ser-197-->Cys-206 loop necessary to achieve this metal 
      coordination. This designed His3Cys metal polyhedron may mimic the zinc binding 
      site in the matrix metalloproteinase prostromelysin.
FAU - Kiefer, L L
AU  - Kiefer LL
AD  - Biochemistry Department, Duke University Medical Center, Durham, North Carolina 
      27710.
FAU - Krebs, J F
AU  - Krebs JF
FAU - Paterno, S A
AU  - Paterno SA
FAU - Fierke, C A
AU  - Fierke CA
LA  - eng
GR  - GM40602/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Isoenzymes)
RN  - 0 (Ligands)
RN  - 0 (Recombinant Proteins)
RN  - 2ZD004190S (Threonine)
RN  - 3G0H8C9362 (Cobalt)
RN  - EC 4.2.1.1 (Carbonic Anhydrases)
RN  - J41CSQ7QDS (Zinc)
RN  - K848JZ4886 (Cysteine)
RN  - O3FX965V0I (Acetazolamide)
SB  - IM
MH  - Acetazolamide/pharmacology
MH  - Base Sequence
MH  - Binding Sites
MH  - Carbonic Anhydrases/*chemistry/genetics/*metabolism
MH  - Cobalt/metabolism
MH  - *Cysteine
MH  - Humans
MH  - Hydrogen Bonding
MH  - Isoenzymes/chemistry/genetics/metabolism
MH  - Kinetics
MH  - Ligands
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Protein Engineering
MH  - Recombinant Proteins/chemistry/metabolism
MH  - Spectrophotometry/methods
MH  - *Threonine
MH  - Zinc/*metabolism
EDAT- 1993/09/28 00:00
MHDA- 1993/09/28 00:01
CRDT- 1993/09/28 00:00
PHST- 1993/09/28 00:00 [pubmed]
PHST- 1993/09/28 00:01 [medline]
PHST- 1993/09/28 00:00 [entrez]
AID - 10.1021/bi00089a004 [doi]
PST - ppublish
SO  - Biochemistry. 1993 Sep 28;32(38):9896-900. doi: 10.1021/bi00089a004.