Phosphorylation of mitochondrial proteins in isolated porcine ovarian follicles after treatment with luteinizing hormone
LH treatment of isolated medium (3- to 7-mm) follicles from porcine ovaries stimulated pregnenolone synthesis by mitochondria isolated from these tissues. This LH stimulation of pregnenolone synthesis was observed regardless of whether exogenous cholesterol was added to the mitochondrial assay syste...
Saved in:
Published in | Endocrinology (Philadelphia) Vol. 114; no. 2; p. 588 |
---|---|
Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
United States
01.02.1984
|
Subjects | |
Online Access | Get more information |
Cover
Loading…
Abstract | LH treatment of isolated medium (3- to 7-mm) follicles from porcine ovaries stimulated pregnenolone synthesis by mitochondria isolated from these tissues. This LH stimulation of pregnenolone synthesis was observed regardless of whether exogenous cholesterol was added to the mitochondrial assay system, suggesting that factors besides cholesterol availability may be important for mitochondrial steroidogenesis. Since ovarian mitochondria contain AMP-dependent protein kinase activity, we investigated the role of protein phosphorylation in regulating mitochondrial steroidogenesis. Mitochondria were isolated from different sized follicles and corpora lutea and incubated for 10 min with [gamma-32P]ATP in the absence or presence of 2 microM cAMP. Mitochondrial proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 32P labeling of mitochondrial proteins was determined by autoradiography. In all mitochondrial preparations from the various ovarian tissues, 32P labeling was found only for proteins with apparent mol wt of 44,000 and 55,000. While cAMP addition was necessary for phosphorylation of the 44,000 mol wt mitochondrial protein(s), phosphorylation of the 55,000 mol wt mitochondrial protein(s) occurred independently of cAMP. Intact medium follicles were preincubated for 120 min in medium 199D containing 32Pi, and then incubated in nonradioactive medium without or with LH (1 microgram/ml) for up to 2 h. Mitochondria were isolated from these follicles, and their proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. While several mitochondrial proteins were labeled in these experiments, only the 44,000 and 55,000 mol wt proteins were studied. After 30 min of incubation, LH treatment stimulated the phosphorylation of a mitochondrial protein with apparent mol wt of 44,000. However, 32P labeling of this 44,000 mol wt mitochondrial protein decreased in LH-treated follicles after incubation for 60 and 120 min. In untreated follicles, 32P labeling of the 44,000 mol wt mitochondrial protein did not change after 30 min of incubation, but phosphorylation did increase after 60- and 120-min incubations. 32P labeling of the 55,000 mol wt mitochondrial protein increased linearly with time in both untreated and LH-treated follicles. Although cAMP treatment of isolated follicular mitochondria was similar to the LH stimulation of intact follicles with respect to phosphorylation of the 44,000 mol wt mitochondrial protein, cAMP treatment of isolated follicular mitochondria did not stimulate pregnenolone synthesis. |
---|---|
AbstractList | LH treatment of isolated medium (3- to 7-mm) follicles from porcine ovaries stimulated pregnenolone synthesis by mitochondria isolated from these tissues. This LH stimulation of pregnenolone synthesis was observed regardless of whether exogenous cholesterol was added to the mitochondrial assay system, suggesting that factors besides cholesterol availability may be important for mitochondrial steroidogenesis. Since ovarian mitochondria contain AMP-dependent protein kinase activity, we investigated the role of protein phosphorylation in regulating mitochondrial steroidogenesis. Mitochondria were isolated from different sized follicles and corpora lutea and incubated for 10 min with [gamma-32P]ATP in the absence or presence of 2 microM cAMP. Mitochondrial proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 32P labeling of mitochondrial proteins was determined by autoradiography. In all mitochondrial preparations from the various ovarian tissues, 32P labeling was found only for proteins with apparent mol wt of 44,000 and 55,000. While cAMP addition was necessary for phosphorylation of the 44,000 mol wt mitochondrial protein(s), phosphorylation of the 55,000 mol wt mitochondrial protein(s) occurred independently of cAMP. Intact medium follicles were preincubated for 120 min in medium 199D containing 32Pi, and then incubated in nonradioactive medium without or with LH (1 microgram/ml) for up to 2 h. Mitochondria were isolated from these follicles, and their proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. While several mitochondrial proteins were labeled in these experiments, only the 44,000 and 55,000 mol wt proteins were studied. After 30 min of incubation, LH treatment stimulated the phosphorylation of a mitochondrial protein with apparent mol wt of 44,000. However, 32P labeling of this 44,000 mol wt mitochondrial protein decreased in LH-treated follicles after incubation for 60 and 120 min. In untreated follicles, 32P labeling of the 44,000 mol wt mitochondrial protein did not change after 30 min of incubation, but phosphorylation did increase after 60- and 120-min incubations. 32P labeling of the 55,000 mol wt mitochondrial protein increased linearly with time in both untreated and LH-treated follicles. Although cAMP treatment of isolated follicular mitochondria was similar to the LH stimulation of intact follicles with respect to phosphorylation of the 44,000 mol wt mitochondrial protein, cAMP treatment of isolated follicular mitochondria did not stimulate pregnenolone synthesis. |
Author | Bieszczad, R R Neymark, M A Dimino, M J |
Author_xml | – sequence: 1 givenname: M A surname: Neymark fullname: Neymark, M A – sequence: 2 givenname: R R surname: Bieszczad fullname: Bieszczad, R R – sequence: 3 givenname: M J surname: Dimino fullname: Dimino, M J |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/6317360$$D View this record in MEDLINE/PubMed |
BookMark | eNotkE1PxCAYhDmsWdfVqzcT_gAKpS1wNBu_kk30oOcNLS8WQ6EBVl1_vTXuaTLJ5JnJnKFFiAEQumT0mlWM3kAwkTBWk4o0Ui7QilLGiagqcYrOcv6YbV3XfImWLWeCt3SFvl-GmKchpoPXxcWAo8WjK7EfYjDJaY-nFAu4kLEL2OU4x8DgKabeBcDxU8-hgG303vUeMta2QMIlgS4jhIK_XBmw3_8h3I8L73juGufZ5-jEap_h4qhr9HZ_97p5JNvnh6fN7Zb0nKlCpG5kp3SnlDXQMFUbaq3WDTTS1Ez3RlDOW6UElVY2jLKuE0aIFtq65VLLao2u_rnTvhvB7KbkRp0Ou-MD1S8Iv2F2 |
CitedBy_id | crossref_primary_10_1016_0014_5793_86_80530_0 |
ContentType | Journal Article |
DBID | CGR CUY CVF ECM EIF NPM |
DOI | 10.1210/endo-114-2-588 |
DatabaseName | Medline MEDLINE MEDLINE (Ovid) MEDLINE MEDLINE PubMed |
DatabaseTitle | MEDLINE Medline Complete MEDLINE with Full Text PubMed MEDLINE (Ovid) |
DatabaseTitleList | MEDLINE |
Database_xml | – sequence: 1 dbid: NPM name: PubMed url: https://proxy.k.utb.cz/login?url=http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed sourceTypes: Index Database – sequence: 2 dbid: EIF name: MEDLINE url: https://proxy.k.utb.cz/login?url=https://www.webofscience.com/wos/medline/basic-search sourceTypes: Index Database |
DeliveryMethod | no_fulltext_linktorsrc |
Discipline | Medicine Anatomy & Physiology |
ExternalDocumentID | 6317360 |
Genre | Research Support, U.S. Gov't, Non-P.H.S Journal Article |
GroupedDBID | --- -DZ -~X .55 .GJ .XZ 08P 0R~ 18M 1TH 29G 2WC 34G 354 39C 3O- 3V. 4.4 48X 53G 5GY 5RE 5RS 5YH 79B 8F7 AABZA AACZT AAIMJ AAJQQ AAKAS AAPGJ AAPQZ AAPXW AARHZ AAUAY AAUQX AAVAP AAWDT AAYJJ ABEFU ABHFT ABJNI ABLJU ABMNT ABNHQ ABPPZ ABPQP ABPTD ABQNK ABSAR ABWST ABXVV ACFRR ACGFO ACGFS ACIPB ACIWK ACPRK ACUTJ ACZBC ADBBV ADGKP ADGZP ADHKW ADIYS ADQBN ADRTK ADVEK ADZCM AELWJ AEMDU AENEX AENZO AETBJ AEWNT AFFNX AFFZL AFGWE AFOFC AFRAH AFULF AFXAL AFYAG AGINJ AGKRT AGMDO AGQXC AGUTN AHMBA AJEEA ALMA_UNASSIGNED_HOLDINGS APIBT APJGH AQKUS ARIXL ATGXG AVNTJ BAWUL BAYMD BCRHZ BENPR BEYMZ BPHCQ BSWAC BTRTY BVXVI C1A C45 CDBKE CGR CJ0 CS3 CUY CVF DAKXR DIK DU5 E3Z EBS ECM EIF EJD EMOBN ENERS F5P FA8 FECEO FHSFR FLUFQ FOEOM FOTVD FQBLK GAUVT GJXCC GX1 H13 HF~ HZ~ H~9 IAO IH2 IHR ITC J5H KBUDW KOP KQ8 KSI KSN L7B LMP M5~ MBLQV MHKGH MJL MVM NLBLG NOMLY NOYVH NPM NVLIB O9- OAUYM OBH ODMLO OFXIZ OHH OHT OJZSN OK1 OPAEJ OVD OVIDX P2P PQQKQ PROAC Q-A REU ROX ROZ TEORI TJX TLC TMA TR2 TWZ UPT VQP VVN W2D W8F WH7 WHG WOQ X52 X7M XJT XOL YBU YHG YOC YQI YSK YYP ZCA ZCG ZGI ZKB ZXP ZY1 |
ID | FETCH-LOGICAL-c319t-8a58b9ab99fde5194d0ffaa5e58d41acd7033699708f85101bb7d776e64638a82 |
ISSN | 0013-7227 |
IngestDate | Sat Sep 28 07:33:22 EDT 2024 |
IsPeerReviewed | true |
IsScholarly | true |
Issue | 2 |
Language | English |
LinkModel | OpenURL |
MergedId | FETCHMERGED-LOGICAL-c319t-8a58b9ab99fde5194d0ffaa5e58d41acd7033699708f85101bb7d776e64638a82 |
PMID | 6317360 |
ParticipantIDs | pubmed_primary_6317360 |
PublicationCentury | 1900 |
PublicationDate | 1984-Feb |
PublicationDateYYYYMMDD | 1984-02-01 |
PublicationDate_xml | – month: 02 year: 1984 text: 1984-Feb |
PublicationDecade | 1980 |
PublicationPlace | United States |
PublicationPlace_xml | – name: United States |
PublicationTitle | Endocrinology (Philadelphia) |
PublicationTitleAlternate | Endocrinology |
PublicationYear | 1984 |
SSID | ssj0014443 |
Score | 1.33192 |
Snippet | LH treatment of isolated medium (3- to 7-mm) follicles from porcine ovaries stimulated pregnenolone synthesis by mitochondria isolated from these tissues. This... |
SourceID | pubmed |
SourceType | Index Database |
StartPage | 588 |
SubjectTerms | Animals Cyclic AMP - pharmacology Female Kinetics Luteinizing Hormone - pharmacology Mitochondria - drug effects Mitochondria - metabolism Ovarian Follicle - drug effects Ovarian Follicle - metabolism Ovarian Follicle - physiology Phosphorylation Protein Kinases - metabolism Proteins - metabolism Swine |
Title | Phosphorylation of mitochondrial proteins in isolated porcine ovarian follicles after treatment with luteinizing hormone |
URI | https://www.ncbi.nlm.nih.gov/pubmed/6317360 |
Volume | 114 |
hasFullText | |
inHoldings | 1 |
isFullTextHit | |
isPrint | |
link | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnV1Rb9MwELY6EGgvCDYmxgD5AfGCjJrEsZ3HCYamSa0Q2qS9TXZtq5VoUo2C1r7yx3dnu0lbxgS8RFUcubHvy-Xu8t0dIW-FyyXYqZrBq8UybvKKGe0qZnXOpfeZKwRmIw-G4vSCn12Wl73erzXW0o-5-TBa3plX8j9ShXMgV8yS_QfJtpPCCfgN8oUjSBiOfyXjL-Pm-2zcXC--tYbfFJ5Q0Gi1Dd04QhWGSeSKT-BeNNqXYHHj1_T3zU_wkzHGgXW5kRyXGoZ33PMQpIUFwBSTJcYU4L9gcRvsoZPaNqB5Ynge7VUM0WDtydkYebhtnGHoFtPEyx6sBVDBUV-OlhFnXzv24idsNtbEi8-6yERWKb4iM3faNiuYzGPuf6ttY85oglW-pjvL2N_vN50OTmmQrm0AUJyB77x5IchkNg0SFmAMFbE9wb2DWxW208gO2ZEKleQQAz7pQxTniXiZlpLqfmLi0-Yd7ZJHaZ4tByUYKudPyZPkYdDjCJdnpOfqPbJ_XOt5M13QdzRwfoO09sjjQaJW7JObLTDRxtMNMNEVmOikpisw0QQmmsBEWzDRACbagokimOgamGgC03Ny8fnk_OMpS2052Aj09ZwpXSpTaVNV3jpwALjte6916UpleaZHFl4ihagq2Vdeoco3RlophRMclL1W-QF5UMP0LwjlmecuU6b0Iuc-K4yQBXdO9GGrC3iTHJKDuJVXs1h75Srt8cs_DRyR3Q6Kr8hDD4-6ew1249y8CZK9BRt1ccY |
link.rule.ids | 786 |
linkProvider | National Library of Medicine |
openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=Phosphorylation+of+mitochondrial+proteins+in+isolated+porcine+ovarian+follicles+after+treatment+with+luteinizing+hormone&rft.jtitle=Endocrinology+%28Philadelphia%29&rft.au=Neymark%2C+M+A&rft.au=Bieszczad%2C+R+R&rft.au=Dimino%2C+M+J&rft.date=1984-02-01&rft.issn=0013-7227&rft.volume=114&rft.issue=2&rft.spage=588&rft_id=info:doi/10.1210%2Fendo-114-2-588&rft_id=info%3Apmid%2F6317360&rft_id=info%3Apmid%2F6317360&rft.externalDocID=6317360 |
thumbnail_l | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/lc.gif&issn=0013-7227&client=summon |
thumbnail_m | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/mc.gif&issn=0013-7227&client=summon |
thumbnail_s | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/sc.gif&issn=0013-7227&client=summon |