Identification by amino acid sequencing of three major regulatory phosphorylation sites on rat acetyl-CoA carboxylase
Abstract
We have examined the sites phosphorylated on acetyl-CoA carboxylase by three protein kinases which have been shown to inactivate the enzyme, i.e. cyclic-AMP-dependent protein kinase, acetyl-CoA carboxylase kinase-2 (ACK2, purified from rat mammary gland) and the AMP-activated protein kinase (formerly called acetyl-CoA carboxylase kinase-3, purified from rat liver). Each protein kinase phosphorylates two out of three sites (termed 1-3) which have been established by amino acid sequencing. The two sites phosphorylated by each kinase can be recovered on separate peptides, TC1 and TC2, derived by combined digestion of the native enzyme by trypsin and chymotrypsin: TC1 = Ser-2Ser(P)-Met-3Ser(P)-Gly-Leu; TC2 = Arg-Met-1Ser(P)-Phe- Cyclic-AMP-dependent protein kinase phosphorylates sites 1 and 2 exclusively, whereas the AMP-activated protein kinase phosphorylates sites 1 and 3, plus at least one other minor site. ACK2 phosphorylates site 1 and, more slowly, an unidentified site(s) within TC1. We have also established the structures of the single major phosphopeptides (T1 and C1 respectively) which are recovered by HPLC after acetyl-CoA carboxylase phosphorylated by cyclic-AMP-dependent protein kinase is digested with trypsin or chymotrypsin alone. T1 is related to TC1, and has the structure: Ser-Ser(P)-Met-Ser-Gly-Leu-His-Leu-Val-Lys. C1 is identical with TC2. We have carried out studies on the correlation of the activity of acetyl-CoA carboxylase with the occupancy of sites 1, 2 and 3 during phosphorylation by each of the three protein kinases. The results suggest that phosphorylation of site 3 is primarily responsible for the large decrease in Vmax produced by the AMP-activated protein kinase, while phosphorylation of site 1 may be primarily responsible for the increase in A0.5 for citrate and more modest depression of Vmax produced by cyclic-AMP-dependent protein kinase and ACK2. Our results emphasize that amino acid sequence information is essential in the unequivocal interpretation of data from phosphopeptide mapping experiments and allow a more complete interpretation of previous data on phosphorylation of acetyl-CoA carboxylase in intact cells. They also open the way to experiments which could establish the physiological roles of these protein kinases in the control of fatty acid synthesis.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Once activated, AMPK redirects metabolism toward increased catabolism or decreased anabolism through phosphorylation of key proteins across multiple pathways, including protein synthesis, lipid homeostasis, glucose metabolism, and mitochondrial homeostasis.
“Once activated, AMPK redirects metabolism toward increased catabolism or decreased anabolism through the phosphorylation of key proteins across multiple pathways, including protein synthesis (e.g., mammalian target of rapamycin complex (1) [2,3], lipid homeostasis (e.g., acetyl coenzyme A carboxylase, ACC) [4,5], glucose metabolism (e.g., phosphofructokinase (2) [6], and mitochondrial homeostasis”
From AMPK, Mitochondrial Function, and Cardiovascular Disease (2020) · cites this paper as [5]
Cites
Carling, D., Zammit, V. A., & Hardie, D. G. (1987) · FEBS Letters · doi:10.1016/0014-5793(87)80292-2 · Cited by 599 · see citing works
Self-citationMolecular aspects of cellular regulation
Cohen, P. (1980) · Elsevier eBooks · Cited by 393 · see citing works
The role of protein phosphorylation in the hormonal control of enzyme activity
Cohen, P. (1985) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1985.tb09121.x · Cited by 379 · see citing works
[6] Catalytic subunit of cAMP-dependent protein kinase
Reimann, E. M., & Beham, R. A. (1983) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/0076-6879(83)99039-0 · Cited by 211 · see citing works
Brownsey, R. W., & Denton, R. M. (1982) · Biochemical Journal · doi:10.1042/bj2020077 · Cited by 117 · see citing works
Hardie, D. G., & Guy, P. S. (1980) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1980.tb04852.x · Cited by 111 · see citing works
Self-citationThe protein phosphatases involved in cellular regulation
Holmes, C. F. B., CAMPBELL, D. G., Caudwell, F. B., et al. (1986) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1986.tb09473.x · Cited by 100 · see citing works
Self-citationHolland, R., Witters, L. A., & Hardie, D. G. (1984) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1984.tb08105.x · Cited by 99 · see citing works
Self-citationWitters, L. A., Tipper, J. P., & Bacon, G. W. (1983) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(20)81941-3 · Cited by 88 · see citing works
Holland, R., & Hardie, D. G. (1985) · FEBS Letters · doi:10.1016/0014-5793(85)80282-9 · Cited by 76 · see citing works
Self-citationTipper, J. P., & Witters, L. A. (1982) · Biochimica et Biophysica Acta (BBA) - General Subjects · doi:10.1016/0304-4165(82)90354-3 · Cited by 76 · see citing works
Guy, P. S., Cohen, P., & Hardie, D. G. (1981) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1981.tb05160.x · Cited by 73 · see citing works
Self-citationBrownsey, R. W., & Hardie, D. G. (1980) · FEBS Letters · doi:10.1016/0014-5793(80)81048-9 · Cited by 51 · see citing works
Self-citationWang, Y. H., Bell, A. W., Hermodson, M. A., & Roach, P. J. (1986) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)75975-4 · Cited by 45 · see citing works
Munday, M. R., & Hardie, D. G. (1984) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1984.tb08237.x · Cited by 43 · see citing works
Self-citationHaystead, T., & Hardie, D. G. (1986) · Biochemical Journal · doi:10.1042/bj2400099 · Cited by 38 · see citing works
Self-citationHolland, R., Hardie, D. G., CLEGG, R. A., et al. (1985) · Biochemical Journal · doi:10.1042/bj2260139 · Cited by 36 · see citing works
Self-citationBrownsey, R. W., Belsham, G. J., & Denton, R. M. (1981) · FEBS Letters · doi:10.1016/0014-5793(81)80123-8 · Cited by 34 · see citing works
Munday, M. R., & Hardie, D. G. (1986) · Biochemical Journal · doi:10.1042/bj2370085 · Cited by 31 · see citing works
Self-citation5 Acetyl-Coenzyme A Carboxylase
Brownsey, R. W., & Denton, R. M. (1987) · The Enzymes · doi:10.1016/s1874-6047(08)60256-5 · Cited by 26 · see citing works
Gairard, A. (1983) · Biochimie · doi:10.1016/s0300-9084(83)80177-1 · Cited by 15 · see citing works
Munday, M. R., Haystead, T., Holland, R., et al. (1986) · Biochemical Society Transactions · doi:10.1042/bst0140559 · Cited by 11 · see citing works
Self-citationCarling, D., & Hardie, D. G. (1986) · Biochemical Society Transactions · doi:10.1042/bst0141076 · Cited by 11 · see citing works
Self-citationCLEGG, R. A., West, D. W., & Aitchison, R. E. D. (1987) · Biochemical Journal · doi:10.1042/bj2410447 · Cited by 9 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Adipocytes promote ovarian cancer metastasis and provide energy for rapid tumor growth
Nieman, K. M., Kenny, H. A., Penicka, C. V., Ladanyi, A., Buell-Gutbrod, R., Zillhardt, M. R., Romero, I. L., Carey, M. S., Mills, G. B., Hotamisligil, G. S., et al. (2011) · Nature Medicine · doi:10.1038/nm.2492 · Cited by 2,279 · see citing works
The AMP‐Activated Protein Kinase: Fuel Gauge of the Mammalian Cell?
Hardie, D. G., & Carling, D. (1997) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1997.00259.x · Cited by 1,317 · see citing works
Self-citationHawley, S. A., Davison, M., Woods, A., Davies, S. P., Beri, R. K., Carling, D., & Hardie, D. G. (1996) · Journal of Biological Chemistry · doi:10.1074/jbc.271.44.27879 · Cited by 1,264 · see citing works
Self-citationFullerton, M. D., Galic, S., Marcinko, K., Sikkema, S., Pulinilkunnil, T., Chen, Z.-P., O'Neill, H. M., Ford, R. J., Palanivel, R., O'Brien, M., et al. (2013) · Nature Medicine · doi:10.1038/nm.3372 · Cited by 893 · see citing works
Self-citationAMP-Activated Protein Kinase – Development of the Energy Sensor Concept
Hardie, D. G., Hawley, S. A., & Scott, J. W. (2006) · The Journal of Physiology · doi:10.1113/jphysiol.2006.108944 · Cited by 813 · see citing works
Self-citationSvensson, R. U., Parker, S. J., Eichner, L. J., Kolar, M. J., Wallace, M., Brun, S. N., Lombardo, P. S., Van Nostrand, J. L., Hutchins, A., Vera, L., et al. (2016) · Nature Medicine · doi:10.1038/nm.4181 · Cited by 536 · see citing works
Sensing of energy and nutrients by AMP-activated protein kinase
Hardie, D. G. (2011) · The American Journal of Clinical Nutrition · doi:10.3945/ajcn.110.001925 · Cited by 372 · see citing works
Self-citationAMPK regulation of fatty acid metabolism and mitochondrial biogenesis: implications for obesity.
O’Neill, H. M., Holloway, G. P., & Steinberg, G. R. (2013) · Molecular and Cellular Endocrinology · doi:10.1016/j.mce.2012.06.019 · Cited by 364 · see citing works
Carling, D., & Grahame Hardie, D. (1989) · Biochimica et Biophysica Acta (BBA) - Molecular Cell Research · doi:10.1016/0167-4889(89)90014-1 · Cited by 296 · see citing works
Self-citationLin, R., Elf, S., Shan, C., Kang, H.-B., Ji, Q., Zhou, L., Hitosugi, T., Zhang, L., Zhang, S., Seo, J. H., et al. (2015) · Nature Cell Biology · doi:10.1038/ncb3255 · Cited by 287 · see citing works
AMPK, Mitochondrial Function, and Cardiovascular Disease
Wu, S., & Zou, M.-H. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21144987 · Cited by 286 · see citing works
DAVIES, S. P., SIM, A. T. R., & HARDIE, D. G. (1990) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1990.tb15293.x · Cited by 274 · see citing works
Self-citationAMP-activated protein kinase: maintaining energy homeostasis at the cellular and whole-body levels
Hardie, D. G. (2014) · Annual Review of Nutrition · doi:10.1146/annurev-nutr-071812-161148 · Cited by 281 · see citing works
Self-citationThe orphan nuclear receptor Nur77 regulates LKB1 localization and activates AMPK
Zhan, Y.-Y., Chen, Y., Zhang, Q., Zhuang, J.-J., Tian, M., Chen, H.-Z., Zhang, L.-R., Zhang, H.-K., He, J.-P., Wang, W.-J., et al. (2012) · Nature Chemical Biology · doi:10.1038/nchembio.1069 · Cited by 180 · see citing works
Protein kinase B activity is required for the effects of insulin on lipid metabolism in adipocytes
Berggreen, C., Gormand, A., Omar, B., Degerman, E., & Göransson, O. (2009) · American Journal of Physiology-Endocrinology and Metabolism · doi:10.1152/ajpendo.90596.2008 · Cited by 118 · see citing works
- Authors
- MUNDAY, M. R., CAMPBELL, D. G., CARLING, D., & HARDIE, D. G.
- Affiliations
- University of Dundee (GB)
- Venue
- European Journal of Biochemistry
- Publisher
- Wiley
- Published
- 1988-08
- Volume
- 175
- Issue
- 2
- Pages
- 331-338
- ISSN
- 0014-2956 (ISSN-L)
- 1432-1033
- OpenAlex
- W2019677361
- Cited by
- 308
- References
- 24
- Citations by year
- 2026: 14 · 2025: 6 · 2024: 12 · 2023: 9 · 2022: 17 · 2021: 20 · 2020: 13 · 2019: 12 · 2018: 14 · 2017: 9 · 2016: 15 · 2015: 12 · 2014: 14 · 2013: 10 · 2012: 11
- FWCI
- 3.56
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- European Journal of Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- UCL Discovery (University College London) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolism, Diabetes, and Cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Lipids, and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Keywords
- acetyl-CoA carboxylase, phosphorylation sites, protein kinase A, AMPK, enzyme regulation, phosphopeptide mapping, citrate inhibition, enzyme kinetics, rat liver, mammary gland, protein kinases, serine phosphorylation
- MeSH terms
- Acetyl-CoA Carboxylase, Amino Acid Sequence, Animals, Binding Sites, Chromatography, High Pressure Liquid, Chymotrypsin, Female, Kinetics, Ligases, Mammary Glands, Animal, Peptide Fragments, Phosphopeptides, Phosphorylation, Protein Kinases, Trypsin, Rats
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
