PMID- 10820026
OWN - NLM
STAT- MEDLINE
DCOM- 20000706
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 39
IP  - 18
DP  - 2000 May 9
TI  - Identification of the carbonic anhydrase II binding site in the Cl(-)/HCO(3)(-) 
      anion exchanger AE1.
PG  - 5527-33
AB  - The human Cl(-)/HCO(3)(-) anion exchanger (AE1) possesses a binding site within 
      its 33 residue carboxyl-terminal region (Ct) for carbonic anhydrase II (CAII). 
      The amino acid sequence comprising this CAII binding site was determined by 
      peptide competition and by testing the ability of truncation and point mutants of 
      the Ct sequence to bind CAII with a sensitive microtiter plate binding assay. A 
      synthetic peptide consisting of the entire 33 residues of the Ct (residues 
      879-911) could compete with a GST fusion protein of the Ct (GST-Ct) for binding 
      to immobilized CAII, while a peptide consisting of the last 16 residues (896-911) 
      could not. A series of truncation mutants of the GST-Ct showed that the terminal 
      21 residues of AE1 were not required for binding CAII. Removal of four additional 
      residues (887-890) from the Ct resulted in loss of CAII binding. Acidic residues 
      in this region (D887ADD) were critical for binding since mutating this sequence 
      in the GST-Ct to DAAA, AAAA, or NANN caused loss of CAII binding. A GST-Ct 
      construct mutated to D887ANE, the homologous sequence in AE2, could bind CAII. 
      AE2 is a widely expressed anion exchanger and has a homologous Ct region with 60% 
      sequence identity to AE1. A GST fusion protein of the 33 residue Ct of AE2 could 
      bind to CAII similarly to the Ct of AE1. Tethering of CAII to an acidic motif 
      within the Ct of anion exchangers may be a general mechanism for promoting 
      bicarbonate transport across cell membranes.
FAU - Vince, J W
AU  - Vince JW
AD  - Medical Research Council Group in Membrane Biology, Departments of Medicine and 
      Biochemistry, Room 7344, Medical Sciences Building, University of Toronto, 
      Toronto, Canada M5S 1A8.
FAU - Reithmeier, R A
AU  - Reithmeier RA
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Anion Transport Proteins)
RN  - 0 (Antiporters)
RN  - 0 (Chloride-Bicarbonate Antiporters)
RN  - 0 (Enzymes, Immobilized)
RN  - 0 (Membrane Proteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 0 (SLC4A Proteins)
RN  - EC 4.2.1.1 (Carbonic Anhydrases)
SB  - IM
MH  - Amino Acid Sequence
MH  - *Anion Transport Proteins
MH  - Antiporters/*chemistry/genetics
MH  - Binding Sites
MH  - Binding, Competitive
MH  - Biological Transport/genetics
MH  - Carbonic Anhydrases/*chemistry
MH  - Chloride-Bicarbonate Antiporters
MH  - Enzymes, Immobilized
MH  - Humans
MH  - Membrane Proteins/metabolism
MH  - Molecular Sequence Data
MH  - Mutation
MH  - Peptide Fragments/metabolism
MH  - Phosphorylation
MH  - Recombinant Proteins/metabolism
MH  - SLC4A Proteins
MH  - Sequence Alignment
EDAT- 2000/05/23 09:00
MHDA- 2000/07/08 11:00
CRDT- 2000/05/23 09:00
PHST- 2000/05/23 09:00 [pubmed]
PHST- 2000/07/08 11:00 [medline]
PHST- 2000/05/23 09:00 [entrez]
AID - bi992564p [pii]
AID - 10.1021/bi992564p [doi]
PST - ppublish
SO  - Biochemistry. 2000 May 9;39(18):5527-33. doi: 10.1021/bi992564p.