IDENTIFICATION OF THE PLASMA-MEMBRANE PROTEOLIPID PROTEIN AS A CONSTITUENT OF BRAIN COATED VESICLES AND SYNAPTIC PLASMA-MEMBRANE

被引:17
作者
SAPIRSTEIN, VS
NOLAN, C
STERN, R
CIOCCI, M
MASUR, SK
机构
[1] NATHAN KLINE INST, DIV MOLEC BIOL, ORANGEBURG, NY 10962 USA
[2] NYU, FACHBEREICH MED, DEPT PSYCHIAT, NEW YORK, NY 10003 USA
[3] CUNY MT SINAI SCH MED, DEPT PHYSIOL & BIOPHYS, NEW YORK, NY 10029 USA
关键词
D O I
10.1111/j.1471-4159.1988.tb01829.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have analyzed brain coated vesicles and synaptic plasma membrane for the presence of the plasma membrane proteolipid protein. Coated vesicles were isolated from calf brain gray matter with a final purification on Sephacryl S-1000 and reisolated twice by chromatography to ensure homogeneity. Fractions were analyzed by gel electrophoresis, immunoblotting for clathrin heavy chain, and by electron microscopy. Using an immunoblotting assay we were able to demonstrate the presence of the plasma membrane proteolipid protein in these coated vesicles at a significant level (i.e., approximately 1% of the bilayer protein of these vesicles). Reisolation of coated vesicles did not diminish the concentration of the protein in this fraction. Removel of the clathrin coat proteins or exposure of the coated vesicles to 0.1 M Na2CO3 showed that the plasma membrane proteolipid protein is not removed during uncoating and lysis but is intrinsic to the membrane bilayer of these vesicles. These studies demonstrate that plasma membrane proteolipid protein represents a significant amount of the bilayer protein of coated vesicles, suggesting that these vesicles may be a transport vehicle for the intracellular movement of the plasma membrane proteolipid protein. Isolation of synaptic plasma membranes from adult rat brain and estimation of the plasma membrane proteolipid protein content using the immunoblotting method confirmed earlier studies that show this protein is present in this membrane fraction at high levels as well (approximately 1-2%). The levle of this protein in the synaptic plasma membrane suggests that the synaptic plasma membrane is one major site to which these vesicles may be targeted or from which the protein is being retrieved.
引用
收藏
页码:925 / 933
页数:9
相关论文
共 30 条
[1]   DISTRIBUTION OF MEMBRANE-GLYCOPROTEINS AMONG THE ORGANELLES OF A SINGLE IDENTIFIED NEURON OF APLYSIA .2. ISOLATION AND CHARACTERIZATION OF A GLYCOPROTEIN ASSOCIATED WITH VESICLES [J].
AMBRON, RT ;
SHERBANY, AA ;
SCHWARTZ, JH .
BRAIN RESEARCH, 1981, 207 (01) :33-48
[2]   Role of the coated endocytic vesicle in the uptake of receptor-bound low density lipoprotein in human fibroblasts [J].
ANDERSON, RGW ;
BROWN, MS ;
GOLDSTEIN, JL .
CELL, 1977, 10 (03) :351-364
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   COATED PITS ACT AS MOLECULAR FILTERS [J].
BRETSCHER, MS ;
THOMSON, JN ;
PEARSE, BMF .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1980, 77 (07) :4156-4159
[5]   IDENTIFICATION OF A PROTEIN-KINASE AS AN INTRINSIC COMPONENT OF RAT-LIVER COATED VESICLES [J].
CAMPBELL, C ;
SQUICCIARINI, J ;
SHIA, M ;
PILCH, PF ;
FINE, RE .
BIOCHEMISTRY, 1984, 23 (19) :4420-4426
[6]  
CHENG TPO, 1982, BRAIN RES, V239, P201
[7]   SYNAPTIC PLASMA MEMBRANES FROM RAT BRAIN SYNAPTOSOMES - ISOLATION AND PARTIAL CHARACTERIZATION [J].
COTMAN, CW ;
MATTHEWS, DA .
BIOCHIMICA ET BIOPHYSICA ACTA, 1971, 249 (02) :380-+
[8]  
FISCHER I, 1986, J NEUROCHEM, V47, P232
[9]  
FOLCH J, 1951, J BIOL CHEM, V191, P807
[10]   EVIDENCE THAT OUABAIN BINDS TO THE SAME LARGE POLYPEPTIDE-CHAIN OF DIMERIC NA,K-ATPASE THAT IS PHOSPHORYLATED FROM PI [J].
FORBUSH, B ;
HOFFMAN, JF .
BIOCHEMISTRY, 1979, 18 (11) :2308-2315