Effects of sulfhydryl reagents on human platelet membrane proteins
Sulfhydryl (SH)- containing human platelet membrane proteins were studied by treating isolated membranes prepared by nitrogen cavitation technique with azodicarboxylic acid bis(dimethylamide) (DA), 5,5'-dithobis(2-nitrobenzoic) acid (DNTB) and N-ethylmaleimide (NEM). Membrane proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). DA and DTNB, membrane penetrating and non-penetrating reagents respectively, cross-linked the myosin heavy chain into a heavy molecular weight complex (HMWC), providing evidence that the membrane vesicles were inside-out or open. DTNB-treated membranes containing an additional 66K da protein band, not observed in DA-treated or untreated membrane proteins. Pretreatment with NEM prevented myosin cross-linking. Reaction of DA cross-linked membranes with (/sup 3/H)NEM yielded three radiolabeled bands: The HMWC containing myosin, actin and a 24.5 K da protein. Evidence will be presented that the 24.5 K da polypeptide is the protein whose SH alkylation was previously demonstrated to inhibit mobilization of arachidonic acid from platelet membranes.
- Research Organization:
- New York College of Osteopathic Medicine, Old Westbury
- OSTI ID:
- 6374701
- Report Number(s):
- CONF-870644-; TRN: 87-033973
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987
- Country of Publication:
- United States
- Language:
- English
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PROTEINS
CROSS-LINKING
MOLECULAR STRUCTURE
REAGENTS
BIOLOGICAL FUNCTIONS
BLOOD PLATELETS
CELL MEMBRANES
ELECTROPHORESIS
MAN
NEM
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ANIMALS
ANTIMITOTIC DRUGS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CELL CONSTITUENTS
CHEMICAL REACTIONS
DRUGS
FUNCTIONS
IMIDES
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
MATERIALS
MEMBRANES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
POLYMERIZATION
PRIMATES
RADIOSENSITIZERS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques