Aspartate 112 is the selectivity filter of the human voltage-gated proton channel.
The ion selectivity of pumps and channels is central to their ability to perform a multitude of functions. Here we investigate the mechanism of the extraordinary selectivity of the human voltage-gated proton channel, H(V)1 (also known as HVCN1). This selectivity is essential to its ability to regulate reactive oxygen species production by leukocytes, histamine secretion by basophils, sperm capacitation, and airway pH. The most selective ion channel known, H(V)1 shows no detectable permeability to other ions. Opposing classes of selectivity mechanisms postulate that (1) a titratable amino acid residue in the permeation pathway imparts proton selectivity, or (2) water molecules 'frozen' in a narrow pore conduct protons while excluding other ions. Here we identify aspartate 112 as a crucial component of the selectivity filter of H(V)1. When a neutral amino acid replaced Asp 112, the mutant channel lost proton specificity and became anion-selective or did not conduct. Only the glutamate mutant remained proton-specific. Mutation of the nearby Asp 185 did not impair proton selectivity, indicating that Asp 112 has a unique role. Although histidine shuttles protons in other proteins, when histidine or lysine replaced Asp 112, the mutant channel was still anion-permeable. Evidently, the proton specificity of H(V)1 requires an acidic group at the selectivity filter.
Department of Molecular Biophysics & Physiology, Rush University Medical Center, Chicago, Illinois 60612, USA.
Boris
B
Musset
Susan M E
SM
Smith
Sindhu
S
Rajan
Deri
D
Morgan
Vladimir V
VV
Cherny
Thomas E
TE
Decoursey
D001224:Aspartic Acid / Q000235:genetics / Q000378:metabolism*; D004553:Electric Conductivity; D006639:Histidine / Q000235:genetics; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects / Q000235:genetics*; D007473:Ion Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D007552:Isotonic Solutions / Q000494:pharmacology; D008239:Lysine / Q000235:genetics; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D016366:Open Reading Frames / Q000235:genetics; D009994:Osmolar Concentration; D010539:Permeability / Q000187:drug effects; D011522:Protons; D013379:Substrate Specificity / Q000187:drug effects; D013395:Sucrose / Q000494:pharmacology
C510235:HVCN1 protein, human; D007473:Ion Channels; D007552:Isotonic Solutions; D050505:Mutant Proteins; D011522:Protons; D001224:Aspartic Acid; D006639:Histidine; D013395:Sucrose; D008239:Lysine