Band 3 structure and function: sup 35 Cl NMR and topographical investigations

Thesis/Dissertation · Sun Jan 01 04:00:00 EST 1989

OSTI ID:5614511

Kanes, K J

Band 3 is the anion exchange protein in red blood cells. Physiologically, it transports Cl{sup {minus}} into or out of the red blood cells and then transports HCO{sub 3}{sup {minus}} in the opposite direction so that electroneutrality is maintained on both sides of the membrane. The anion exchange mechanism of band 3 is unique among the ion transport proteins. It transports anions by a ping-pong mechanism, meaning it is a gated protein which effects the one-for-one exchange of anions across the membrane. It is also unusual because it transports a wide variety of anions in a very efficient manner (up to 200 sec{sup {minus}1}). An arginine has been implicated in the binding and transport of chloride across the red blood cell membrane. The primary goal of this work was to determine the location of that arginine. A second goal was to investigate divalent anion binding to the active site. {sup 35}Cl NMR was used to investigate the competition of chloride with divalent anions at the chloride binding site of band 3. These studies were performed to determine if divalent anions compete with chloride for binding at the active site. pH profiles of chloride competition with divalent anions were also obtained. The goal of the biochemical studies was to determine the location of the band 3 arginine anion binding site. In order to insure that an arginine preferentially labeled by {sup 14}C-phenylglyoxal was indeed at a transmembrane peptide and to verify the model, many of the transmembrane sequences have been isolated. This was accomplished by modification of a technique developed to separate hydrophobic synthetic peptides. Of a probable total of 14 transmembrane helices, the N-termini and/or C-termini have been determined for 6 of them.

• Research Organization: California Inst. of Tech., Pasadena, CA (United States)
• OSTI ID: 5614511
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
BIOCHEMICAL REACTION KINETICS
BIOCHEMISTRY
BIOLOGICAL FUNCTIONS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBON 14 COMPOUNDS
CELL CONSTITUENTS
CELL MEMBRANES
CHEMISTRY
CHLORIDES
CHLORINE 35
CHLORINE COMPOUNDS
CHLORINE ISOTOPES
ERYTHROCYTES
HALIDES
HALOGEN COMPOUNDS
ION EXCHANGE
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
LIGHT NUCLEI
MAGNETIC RESONANCE
MATERIALS
MEMBRANE PROTEINS
MEMBRANE TRANSPORT
MEMBRANES
MOLECULAR STRUCTURE
NUCLEAR MAGNETIC RESONANCE
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PROTEINS
REACTION KINETICS
RECEPTORS
RESONANCE
STABLE ISOTOPES
TRACER TECHNIQUES