Two Subunits of Glycosylphosphatidylinositol Transamidase, GPI8 and PIG-T, Form a Functionally Important Intermolecular Disulfide Bridge
Open accessAbstract
Many eukaryotic proteins are tethered to the plasma membrane via glycosylphosphatidylinositol (GPI). GPI transamidase is localized in the endoplasmic reticulum and mediates post-translational transfer of preformed GPI to proteins bearing a carboxyl-terminal GPI attachment signal. Mammalian GPI transamidase is a multimeric complex consisting of at least five subunits. Here we report that two subunits of mammalian GPI transamidase, GPI8 and PIG-T, form a functionally important disulfide bond between conserved cysteine residues. GPI8 and PIG-T mutants in which relevant cysteines were replaced with serines were unable to fully restore the surface expression of GPI-anchored proteins upon transfection into their respective mutant cells. Microsomal membranes of these transfectants had markedly decreased activities in an in vitro transamidase assay. The formation of this disulfide bond is not essential but required for full transamidase activity. Antibodies against GPI8 and PIG-T revealed that endogenous as well as exogenous proteins formed a disulfide bond. Furthermore trypanosome GPI8 forms a similar intermolecular disulfide bond via its conserved cysteine residue, suggesting that the trypanosome GPI transamidase is also a multimeric complex likely containing the orthologue of PIG-T. We also demonstrate that an inactive human GPI transamidase complex that consists of non-functional GPI8 and four other components was co-purified with the proform of substrate proteins, indicating that these five components are sufficient to hold the substrate proteins.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
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?
The disulfide bond between Gpi8 and Gpi16 was previously characterized for human GPI-T.
“previously characterized for human GPI-T (94)”
From Computed structures of core eukaryotic protein complexes (2021) · cites this paper as [94]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Cosman, D. J., Müllberg, J., Sutherland, C. L., Chin, W. A., Armitage, R. J., Fanslow, W. C., Kubin, M. Z., & Chalupny, N. J. (2001) · Immunity · doi:10.1016/s1074-7613(01)00095-4 · Cited by 1,228 · see citing works
Takeda, J., Miyata, T., Kawagoe, K., Iida, Y., Endo, Y., Fujita, T., Takahashi, M., Kitani, T., & Kinoshita, T. (1993) · Cell · doi:10.1016/0092-8674(93)90250-t · Cited by 1,059 · see citing works
Self-citationHOW GLYCOSYL-PHOSPHATIDYLINOSITOL-ANCHORED MEMBRANE PROTEINS ARE MADE
Udenfriend, S., & Kodukula, K. (1995) · Annual Review of Biochemistry · doi:10.1146/annurev.bi.64.070195.003023 · Cited by 481 · see citing works
Glycoproteins form mixed disulphides with oxidoreductases during folding in living cells
Molinari, M., & Helenius, A. (1999) · Nature · doi:10.1038/47062 · Cited by 320 · see citing works
Tarutani, M., Itami, S., Okabe, M., Ikawa, M., Tezuka, T., Yoshikawa, K., Kinoshita, T., & Takeda, J. (1997) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.94.14.7400 · Cited by 263 · see citing works
Self-citationKawagoe, K., Kitamura, D., Okabe, M., Taniuchi, I., Ikawa, M., Watanabe, T., Kinoshita, T., & Takeda, J. (1996) · Blood · doi:10.1182/blood.v87.9.3600.bloodjournal8793600 · Cited by 229 · see citing works
Self-citationDissecting and manipulating the pathway for glycosylphos-phatidylinositol-anchor biosynthesis
Kinoshita, T., & Inoue, N. (2000) · Current Opinion in Chemical Biology · doi:10.1016/s1367-5931(00)00151-4 · Cited by 190 · see citing works
Self-citationEisenhaber, B., Peer Bork, B., & Eisenhaber, F. (2001) · Protein Engineering Design and Selection · doi:10.1093/protein/14.1.17 · Cited by 181 · see citing works
PIG‐S and PIG‐T, essential for GPI anchor attachment to proteins, form a complex with GAA1 and GPI8
Ohishi, K., Inoue, N., & Kinoshita, T. (2001) · The EMBO Journal · doi:10.1093/emboj/20.15.4088 · Cited by 177 · see citing works
Self-citationYeast Gpi8p is essential for GPI anchor attachment onto proteins.
Benghezal, M., Benachour, A., Rusconi, S., Aebi, M., & Conzelmann, A. (1996) · The EMBO Journal · doi:10.1002/j.1460-2075.1996.tb01048.x · Cited by 176 · see citing works
Critical roles of glycosylphosphatidylinositol for Trypanosoma brucei
Nagamune, K., Nozaki, T., Maeda, Y., Ohishi, K., Fukuma, T., Hara, T., Schwarz, R. Τ., Sütterlin, C., Brun, R., Riezman, H., et al. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.180230697 · Cited by 173 · see citing works
Self-citationNozaki, M., Ohishi, K., Yamada, N., Kinoshita, T., Nagy, A., & Takeda, J. (1999) · PubMed · Cited by 167 · see citing works
Self-citationAbe, Y., Shirane, K., Yokosawa, H., et al. (1993) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)53726-1 · Cited by 137 · see citing works
Ohishi, K., Inoue, N., Maeda, Y., Takeda, J., Riezman, H., & Kinoshita, T. (2000) · Molecular Biology of the Cell · doi:10.1091/mbc.11.5.1523 · Cited by 126 · see citing works
Self-citationHong, Y., Ohishi, K., Kang, J. Y., Tanaka, S., Inoue, N., Nishimura, J.-I., Maeda, Y., & Kinoshita, T. (2003) · Molecular Biology of the Cell · doi:10.1091/mbc.e02-12-0794 · Cited by 121 · see citing works
Self-citationLeidich, S. D., Kostova, Z., Latek, R. R., Costello, L. C., Drapp, D. A., Gray, W. M., Fassler, J. S., & Orlean, P. (1995) · Journal of Biological Chemistry · doi:10.1074/jbc.270.22.13029 · Cited by 120 · see citing works
Moran, P., & Caras, I. W. (1994) · The Journal of Cell Biology · doi:10.1083/jcb.125.2.333 · Cited by 115 · see citing works
Lillico, S. G., Field, M. C., Blundell, P. A., Coombs, G. H., & Mottram, J. C. (2003) · Molecular Biology of the Cell · doi:10.1091/mbc.e02-03-0167 · Cited by 114 · see citing works
The GPI Transamidase Complex of<i>Saccharomyces cerevisiae</i>Contains Gaa1p, Gpi8p, and Gpi16p
Fraering, P., Imhof, I., Meyer, U., Strub, J.-M., van Dorsselaer, A., Vionnet, C., & Conzelmann, A. (2001) · Molecular Biology of the Cell · doi:10.1091/mbc.12.10.3295 · Cited by 111 · see citing works
Meyer, U., Benghezal, M., Imhof, I., & Conzelmann, A. (2000) · Biochemistry · doi:10.1021/bi992186o · Cited by 87 · see citing works
An Unconventional Role for Cytoplasmic Disulfide Bonds in Vaccinia Virus Proteins
Locker, J. K., & Griffiths, G. W. (1999) · The Journal of Cell Biology · doi:10.1083/jcb.144.2.267 · Cited by 87 · see citing works
Bijlmakers, M.-J., Benaroch, P., & Ploegh, H. L. (1994) · The EMBO Journal · doi:10.1002/j.1460-2075.1994.tb06560.x · Cited by 87 · see citing works
Hilley, J. D., Zawadzki, J. L., McConville, M. J., Coombs, G. H., & Mottram, J. C. (2000) · Molecular Biology of the Cell · doi:10.1091/mbc.11.4.1183 · Cited by 83 · see citing works
Yu, J., Nagarajan, S., Knez, J. J., Udenfriend, S., Chen, R., & Medof, M. E. (1997) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.94.23.12580 · Cited by 77 · see citing works
A Cell-free Assay for Glycosylphosphatidylinositol Anchoring in African Trypanosomes
Sharma, D. K., Vidugirienė, J., Bangs, J. D., & Menon, A. K. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.23.16479 · Cited by 66 · see citing works
Glycosylphosphatidylinositol biosynthesis validated as a drug target for African sleeping sickness
Ferguson, M. A. J. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.97.20.10673 · Cited by 65 · see citing works
Biosynthesis of phosphatidylinositol glycan-anchored membrane proteins.
Kodukula, K., Micanovic, R., Gerber, L., Tamburrini, M., Brink, L., & Udenfriend, S. (1991) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(20)64345-9 · Cited by 59 · see citing works
Evidence for a role for the Dictyostelium Rap1 in cell viability and the response to osmotic stress
Kang, R., Kae, H., Ip, H., Spiegelman, G. B., & Weeks, G. (2002) · Journal of Cell Science · doi:10.1242/jcs.00039 · Cited by 56 · see citing works
Vainauskas, S., Maeda, Y., Kurniawan, H., Kinoshita, T., & Menon, A. K. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m205402200 · Cited by 47 · see citing works
Self-citationKang, X., Szallies, A., Rawer, M., Echner, H., & Duszenko, M. (2002) · Journal of Cell Science · doi:10.1242/jcs.115.12.2529 · Cited by 41 · see citing works
Endoplasmic reticulum proteins involved in glycosylphosphatidylinositol‐anchor attachment
Vidugirienė, J., Vainauskas, S., Johnson, A. E., & Menon, A. K. (2001) · European Journal of Biochemistry · doi:10.1046/j.1432-1327.2001.02106.x · Cited by 39 · see citing works
Early Events in Glycosylphosphatidylinositol Anchor Addition
Spurway, T. D., Dalley, J. A., High, S., & Bulleid, N. J. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m010128200 · Cited by 37 · see citing works
Sharma, D. K., Hilley, J. D., Bangs, J. D., Coombs, G. H., Mottram, J. C., & Menon, A. K. (2000) · Biochemical Journal · doi:10.1042/bj3510717 · Cited by 34 · see citing works
Soluble constituents of the ER lumen are required for GPI anchoring of a model protein.
Vidugirienė, J., & Menon, A. K. (1995) · The EMBO Journal · doi:10.1002/j.1460-2075.1995.tb00150.x · Cited by 26 · see citing works
Patnaik, P. K., Fang, X., & Cross, G. (1994) · Nucleic Acids Research · doi:10.1093/nar/22.20.4111 · Cited by 23 · see citing works
Flagged first, then by citations
Research works printed but not linked: 4
- Tarutani, M., Itami, S., Okabe, M., Ikawa, M., Tezuka, T., Yoshikawa, K., Kinoshita, T., and Takeda, J. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 7400 -7405
- Nagamune, K., Nozaki, T., Maeda, Y., Ohishi, K., Fukuma, T., Hara, T., Schwarz, R. T., Sutterlin, C., Brun, R., Riezman, H., and Kinoshita, T. (2000) Proc. Natl. Acad. Sci. U. S. A. 97, 10336 -10341
- Ferguson, M. A. J. (2000) Proc. Natl. Acad. Sci. U. S. A. 97, 10673-10675
- Yu, J., Nagarajan, S., Knez, J. J., Udenfriend, S., Chen, R., and Medof, M. E. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 12580 -12585
Cited by
Sorted by citations
Paroxysmal nocturnal hemoglobinuria
Brodsky, R. A. (2014) · Blood · doi:10.1182/blood-2014-02-522128 · Cited by 648 · see citing works
Computed structures of core eukaryotic protein complexes
Humphreys, I. R., Pei, J., Baek, M., Krishnakumar, A., Anishchenko, I., Ovchinnikov, S., Zhang, J., Ness, T. J., Banjade, S., Bagde, S. R., et al. (2021) · Science · doi:10.1126/science.abm4805 · Cited by 542 · see citing works
GPI Transamidase and GPI anchored proteins: Oncogenes and biomarkers for cancer
Gamage, D. G., & Hendrickson, T. L. (2013) · Critical Reviews in Biochemistry and Molecular Biology · doi:10.3109/10409238.2013.831024 · Cited by 63 · see citing works
Vainauskas, S., & Menon, A. K. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m312191200 · Cited by 34 · see citing works
Meitzler, J. L., Gray, J. J., & Hendrickson, T. L. (2007) · Archives of Biochemistry and Biophysics · doi:10.1016/j.abb.2007.03.035 · Cited by 18 · see citing works
Yi, L., Bozkurt, G., Li, Q., Lo, S., Menon, A. K., & Wu, H. (2017) · Scientific Reports · doi:10.1038/srep45912 · Cited by 12 · see citing works
Gamage, D. G., Varma, Y., Meitzler, J. L., Morissette, R., Ness, T. J., & Hendrickson, T. L. (2017) · Archives of Biochemistry and Biophysics · doi:10.1016/j.abb.2017.09.006 · Cited by 8 · see citing works
- Authors
- Ohishi, K., Nagamune, K., Maeda, Y., & Kinoshita, T.
- Affiliations
- Research Institute for Microbial Diseases, The University of Osaka (JP)
- Corresponding author
- Kinoshita, Taroh
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2003-04
- Volume
- 278
- Issue
- 16
- Pages
- 13959-13967
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2041257080
- Cited by
- 83
- References
- 35
- Citations by year
- 2025: 2 · 2024: 5 · 2023: 4 · 2022: 4 · 2021: 9 · 2020: 10 · 2019: 3 · 2018: 2 · 2017: 4 · 2016: 1 · 2015: 2 · 2014: 5 · 2013: 5 · 2012: 1
- FWCI
- 1.15
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Ministry of Education, Culture, Sports, Science and Technology
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Lipid Membrane Structure and Behavior
- Keywords
- intermolecular disulfide bonds, cysteine residues, GPI-anchored proteins, endoplasmic reticulum
- MeSH terms
- Acyltransferases, Animals, Cysteine, Disulfides, Flow Cytometry, Cricetinae, Humans, Mutation, Transfection, Protein Biosynthesis, Trypanosoma brucei brucei, Cell Adhesion Molecules, CHO Cells, Protein Structure, Tertiary, CD59 Antigens, K562 Cells
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