Hydrophobic photolabeling identifies BHA2 as the subunit mediating the interaction of bromelain-solubilized influenza virus hemagglutinin with liposomes at low pH
To investigate the molecular basis of the low-pH-mediated interaction of the bromelain-solubilized ectodomain of influenza virus hemagglutinin (BHA) with membranes, we have photolabeled BHA in the presence of liposomes with the two carbene-generating, membrane-directed reagents 3-(trifluoromethyl)-3-(m-(/sup 125/I)iodophenyl)diazirine ((/sup 125/I)TID) and a new analogue of a phospholipid, 1-palmitoyl-2-(11-(4-(3-(trifluoromethyl)diazirinyl)phenyl)(2-/sup 3/H) undecanoyl)-sn-glycero-3-phosphocholine ((/sup 3/H)-PTPC/11). With the latter reagent, BHA was labeled in a strictly pH-dependent manner, i.e., at pH 5 only, whereas with (/sup 125/I)TID, labeling was seen also at pH 7. In all experiments, the label was selectively incorporated into the BHA2 polypeptide, demonstrating that the interaction of BHA with membranes is mediated through this subunit, possibly via its hydrophobic N-terminal segment. Similar experiments with a number of other water-soluble proteins (ovalbumin, carbonic anhydrase, alpha-lactalbumin, trypsin, and soybean trypsin inhibitor) indicate that the ability to interact with liposomes at low pH is not a property specific for BHA but is observed with other, perhaps most, proteins.
- Research Organization:
- Laboratorium fuer Biochemie der Eidgenoessischen Technischen Hochschule, Zuerich (Switzerland)
- OSTI ID:
- 6908812
- Journal Information:
- Biochemistry; (United States), Vol. 27:6
- Country of Publication:
- United States
- Language:
- English
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HEMAGGLUTININS
BIOCHEMICAL REACTION KINETICS
LABELLING
CELL MEMBRANES
INFLUENZA VIRUSES
IODINE 125
LIPOSOMES
MOLECULAR BIOLOGY
NUCLEAR MAGNETIC RESONANCE
PH VALUE
REAGENTS
SOLUBILITY
TRACER TECHNIQUES
TRITIUM COMPOUNDS
AGGLUTININS
ANTIBODIES
BETA DECAY RADIOISOTOPES
CELL CONSTITUENTS
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
MAGNETIC RESONANCE
MEMBRANES
MICROORGANISMS
NUCLEI
ODD-EVEN NUCLEI
ORGANOIDS
PARASITES
RADIOISOTOPES
REACTION KINETICS
RESONANCE
VIRUSES
550201* - Biochemistry- Tracer Techniques