Association of lipids with integral membrane surface proteins of Mycoplasma hyorhinis
Triton X-114 (TX-114)-phase fractionation was used to identify and characterize integral membrane surface proteins of the wall-less procaryote Mycoplasma hyorhinis GDL. Phase fractionation of mycoplasmas followed by analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed selective partitioning of approximately 30 (/sup 35/S)methionine-labeled intrinsic membrane proteins into the TX-114 phase. Similar analysis of (/sup 3/H)palmitate-labeled cells showed that approximately 20 proteins of this organism were associated with lipid, all of which also efficiently partitioned as integral membrane components into the detergent phase. Immunoblotting and immunoprecipitation of TX-114-phase proteins from /sup 125/I-surface-labeled cells with four monoclonal antibodies to distinct surface epitopes of M. hyorhinis identified surface proteins p120, p70, p42, and p23 as intrinsic membrane components. Immunoprecipitation of (/sup 3/H)palmitate-labeled TX-114-phase proteins further established that surface proteins p120, p70, and p23 (a molecule that mediates complement-dependent mycoplasmacidal monoclonal antibody activity) were among the lipid-associated proteins of this organism. Two of these proteins, p120 and p123, were acidic (pI less than or equal to 4.5), as shown by two-dimensional isoelectric focusing. This study established that M. hyorhinis contains an abundance of integral membrane proteins tightly associated with lipids and that many of these proteins are exposed at the external surface of the single limiting plasma membrane. Monoclonal antibodies are reported that will allow detailed analysis of the structure and processing of lipid-associated mycoplasma proteins.
- Research Organization:
- Univ. of Missouri-Columbia (USA)
- OSTI ID:
- 5446406
- Journal Information:
- Infect. Immun.; (United States), Vol. 56:2
- Country of Publication:
- United States
- Language:
- English
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MEMBRANE PROTEINS
FRACTIONATION
IMMUNOASSAY
DETERGENTS
ELECTROPHORESIS
HEXADECANOIC ACID
IODINE 125
LIPIDS
METHIONINE
MOLECULAR WEIGHT
MONOCLONAL ANTIBODIES
MYCOPLASMA
SOLUBILITY
SULFUR 35
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ADDITIVES
AMINO ACIDS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARBOXYLIC ACIDS
DAYS LIVING RADIOISOTOPES
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
EMULSIFIERS
EVEN-ODD NUCLEI
INTERMEDIATE MASS NUCLEI
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LIGHT NUCLEI
LIPOTROPIC FACTORS
MICROORGANISMS
MONOCARBOXYLIC ACIDS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PROTEINS
RADIOISOTOPES
SEPARATION PROCESSES
SULFUR ISOTOPES
SURFACTANTS
WETTING AGENTS
550201* - Biochemistry- Tracer Techniques