In vivo trafficking of nascent H+-K+-ATPase in rabbit parietal cells

被引:11
作者
Crothers, JM [1 ]
Chow, DC [1 ]
Scalley, ML [1 ]
Forte, JG [1 ]
机构
[1] UNIV CALIF BERKELEY, DEPT MOLEC & CELL BIOL, DIV CELL & DEV BIOL, BERKELEY, CA 94720 USA
来源
AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY | 1995年 / 269卷 / 06期
关键词
proton pump; posttranslational processing; protein turnover; glycoprotein; acid secretion;
D O I
10.1152/ajpgi.1995.269.6.G883
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
Protein metabolic labeling in vivo was used to determine a time course for trafficking of nascent H+-K+-adenosinetriphosphatase (H+K+-ATPase) from endoplasmic reticulum (ER) to mature tubulovesicles in parietal cells. Stomachs of cimetidine-treated rabbits were taken 15-90 min after injection of [S-35]methionine/cysteine, and mucosal microsomes were fractionated on sucrose gradients for analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blot, and autoradiography. After 15 min, labeled alpha-subunit peaked at similar to 1.14 g/ml, matching the distribution of the high-mannose beta-subunit precursor, ''pre-beta.'' After 30 min, most labeled alpha-subunit was in a peak at similar to 1.10 g/ml, considered to be Golgi. By 90 min, most labeled alpha-subunit was in a light peak, at similar to 1.07 g/ml, aligned with the major peak. of total H+-K+-ATPase previously characterized as mature tubulovesicles. From material enriched in pre-beta, alpha-subunit was coprecipitated with pre-beta by a terminal mannose-specific lectin, Galanthus nivalis agglutinin, in the same ratio as the mature alpha:beta ratio. Thus alpha- and beta-subunits associated early in the ER. This is the first use of protein metabolic labeling to study early trafficking of the H+-K+-ATPase in vivo. The techniques may be usefully applied to examining changes in H+-K+-ATPase synthetic rate in response to various pharmacological treatments and studying the divergent pathways for nascent H+-K+- and Na+-K+-ATPases.
引用
收藏
页码:G883 / G891
页数:9
相关论文
共 31 条
  • [1] MUTUAL DEPENDENCE OF NA,K-ATPASE ALPHA-SUBUNITS AND BETA-SUBUNITS FOR CORRECT POSTTRANSLATIONAL PROCESSING AND INTRACELLULAR-TRANSPORT
    ACKERMANN, U
    GEERING, K
    [J]. FEBS LETTERS, 1990, 269 (01): : 105 - 108
  • [2] ACKERMANN U, 1992, J BIOL CHEM, V267, P12911
  • [3] THE BETA-SUBUNIT OF THE GASTRIC H+/K+-ATPASE CAN OCCUR WITHOUT THE ALPHA-SUBUNIT
    BALDWIN, GS
    [J]. FEBS LETTERS, 1990, 272 (1-2) : 159 - 162
  • [4] IN-SITU HYBRIDIZATION OF MESSENGER-RNA FOR THE GASTRIC H+, K+-ATPASE IN RAT OXYNTIC MUCOSA
    BAMBERG, K
    NYLANDER, S
    HELANDER, KG
    LUNDBERG, LG
    SACHS, G
    HELANDER, HF
    [J]. BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 1994, 1190 (02): : 355 - 359
  • [5] EFFECT OF OMEPRAZOLE ON GENE-EXPRESSION IN CANINE GASTRIC PARIETAL-CELLS
    CAMPBELL, VW
    YAMADA, T
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY, 1991, 260 (03): : G434 - G439
  • [6] CAMPBELL VW, 1989, J BIOL CHEM, V264, P11381
  • [7] CAPLAN MJ, 1990, J BIOL CHEM, V265, P3528
  • [8] CHARACTERIZATION OF THE BETA-SUBUNIT OF THE H+-K+-ATPASE USING AN INHIBITORY MONOCLONAL-ANTIBODY
    CHOW, DC
    FORTE, JG
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY, 1993, 265 (06): : C1562 - C1570
  • [9] GASTRIC H+-K+-ATPASE ACTIVITY IS INHIBITED BY REDUCTION OF DISULFIDE BONDS IN BETA-SUBUNIT
    CHOW, DC
    BROWNING, CM
    FORTE, JG
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY, 1992, 263 (01): : C39 - C46
  • [10] ONTOGENY OF GASTRIC H+-K+-ATPASE IN SUCKLING RABBITS
    CROTHERS, JM
    REENSTRA, WW
    FORTE, JG
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY, 1990, 259 (06): : G913 - G921