1. B.P. Konstantinov Petersburg Institute of Nuclear Physics, National Research Center “Kurchatov Institute”, Gatchina, Leningrad Region, Russia 2. Institute of Biomedical Chemistry, Moscow, Russia
Existing knowledge on changes of the haptoglobin (Hp) molecule suggests that it may exist in multiple proteoforms, which obviously exhibit different functions. Using two-dimensional electrophoresis (2DE) in combination with mass spectrometry and immunodetection, we have analyzed blood plasma samples from both healthy donors and patients with primary grade IV glioblastoma (GBM), and obtained a detailed composite 2DE distribution map of β-chain proteoforms, as well as the full-length form of Hp (zonulin). Although the total level of plasma Hp exceeded normal values in cancer patients (especially patients with GBM), the presence of particuar proteoforms, detected by their position on the 2DE map, was very individual. Variability was found in both zonulin and the Hp β-chain. The presence of an alkaline form of zonulin in plasma can be considered a conditional, but insufficient, GBM biomarker. In other words, we found that at the level of minor proteoforms of Hp, even in normal conditions, there was a high individual variability. On the one hand, this raises questions about the reasons for such variability, if it is present not only in Hp, but also in other proteins. On the other hand, this may explain the discrepancy between the number of experimentally detected proteoforms and the theoretically possible ones not only in Hp, but also in other proteins.
Ronzhina N.L. et al. Variability of haptoglobin beta-chain proteoforms // Biomeditsinskaya Khimiya. - 2024. - V. 70. -N 2. - P. 114-124.
Ronzhina N.L. et al., "Variability of haptoglobin beta-chain proteoforms." Biomeditsinskaya Khimiya 70.2 (2024): 114-124.
Ronzhina, N. L., Zorina, E. S., Zavialova, M. G., Legina, O. K., Naryzhny, S. N. (2024). Variability of haptoglobin beta-chain proteoforms. Biomeditsinskaya Khimiya, 70(2), 114-124.
References
Zhang S., Shang S., Li W., Qin X., Liu Y. (2016) Insights on N-glycosylation of human haptoglobin and its association with cancers. Glycobiology, 26(7), 684-692. CrossRef Scholar google search
Kumar D.M., Thota B., Shinde S.V., Prasanna K.V., Hegde A.S., Arivazhagan A., Chandramouli B.A., Santosh V., Somasundaram K. (2010) Proteomic identification of haptoglobin α2 as a glioblastoma serum biomarker: Implications in cancer cell migration and tumor growth. J. Proteome Res., 9(11), 5557-5567. CrossRef Scholar google search
Gollapalli K., Ray S., Srivastava R., Renu D., Singh P., Dhali S., Bajpai Dikshit J., Srikanth R., Moiyadi A., Srivastava S. (2012) Investigation of serum proteome alterations in human glioblastoma multiforme. Proteomics, 12(14), 2378-2390. CrossRef Scholar google search
Garibay-Cerdenares O.L., Hernández-Ramírez V.I., Osorio-Trujillo J.C., Hernández-Ortíz M., Gallardo-Rincón D., Cantú de León D., Encarnación-Guevara S., Villegas-Pineda J.C., Talamás-Rohana P. (2014) Proteomic identification of fucosylated haptoglobin alpha isoforms in ascitic fluids and its localization in ovarian carcinoma tissues from Mexican patients. J. Ovarian Res., 7(1), 27. CrossRef Scholar google search
Ye B., Cramer D.W., Skates S.J., Gygi S.P., Pratomo V., Fu L., Horick N.K., Licklider L.J., Schorge J.O., Berkowitz R.S., Mok S.C. (2003) Haptoglobin-α subunit as potential serum biomarker in ovarian cancer: Identification and characterization using proteomic profiling and mass spectrometry. Clin. Cancer Res., 9(8), 2904-2911. Scholar google search
Naryzhny S.N., Legina O.K. (2021) Haptoglobin as a biomarker. Biomeditsinskaya Khimiya, 67(2), 105-118. CrossRef Scholar google search
Polonovski M., Jayle M. (1938) Existance dans le plasma sanguin d’une substance activant l’action peroxydasique de l’hemoglobine. C. R. Soc. Biol., 129, 457-460. Scholar google search
Sultan A., Raman B., Rao C.M., Tangirala R. (2013) The extracellular chaperone haptoglobin prevents serum fatty acid-promoted amyloid fibril formation of β2-microglobulin, resistance to lysosomal degradation, and cytotoxicity. J. Biol. Chem., 288(45), 32326-32342. CrossRef Scholar google search
Cohen A.L., Colman H. (2015) Glioma biology and molecular markers. Cancer Treat. Res., 163, 15-30. CrossRef Scholar google search
Shih A.W.Y., Mcfarlane A., Verhovsek M. (2014) Haptoglobin testing in hemolysis: Measurement and interpretation. Am. J. Hematol., 89(4), 443-447. CrossRef Scholar google search
Bowman B.H., Kurosky A. (1982) Haptoglobin: The evolutionary product of duplication, unequal crossing over, and point mutation. Advances in Human Genetics, 12, 189-261, 453-454. CrossRef Scholar google search
Clerc F., Reiding K.R., Jansen B.C., Kammeijer G.S.M., Bondt A., Wuhrer M. (2016) Human plasma protein N-glycosylation. Glycoconjugate J., 33(3), 309-343. CrossRef Scholar google search
Pompach P., Brnakova Z., Sanda M., Wu J., Edwards N., Goldman R. (2013) Site-specific glycoforms of haptoglobin in liver cirrhosis and hepatocellular carcinoma. Mol. Cell. Proteomics, 12, 1281-1293. CrossRef Scholar google search
Mikkat S., Koy C., Ulbrich M., Ringel B., Glocker M.O. (2004) Mass spectrometric protein structure characterization reveals cause of migration differences of haptoglobin α chains in two-dimensional gel electrophoresis. Proteomics, 4(12), 3921-3932. CrossRef Scholar google search
Fasano A. (2011) Zonulin and its regulation of intestinal barrier function: The biological door to inflammation, autoimmunity, and cancer. Physiol. Rev., 91(1), 151-175. CrossRef Scholar google search
Naryzhny S., Ronzhina N., Zorina E., Kabachenko F., Zavialova M., Zgoda V., Klopov N., Legina O., Pantina R. (2021) Evaluation of haptoglobin and its proteoforms as glioblastoma markers. Int. J. Mol. Sci., 22(12), 1-12. CrossRef Scholar google search
Naryzhny S.N., Maynskova M.A., Zgoda V.G., Ronzhina N.L., Kleyst O.A., Vakhrushev I.V., Archakov A.I. (2016) Virtual-experimental 2DE approach in chromosome-centric human proteome project. J. Proteome Res., 15(2), 525-530. CrossRef Scholar google search
Hoogland C., Mostaguir K., Sanchez J.C., Hochstrasser D.F., Appel R.D. (2004) SWISS-2DPAGE, ten years later. Proteomics, 4, 2352-2356. CrossRef Scholar google search
Naryzhny S.N., Ronzhina N.L., Mainskova M.A., Belyakova N.V., Pantina R.A., Filatov M.V. (2014) Development of barcode and proteome profiling of glioblastoma. Biomeditsinskaya Khimiya, 60(3), 308-321. CrossRef Scholar google search
Petrenko E.S., Kopylov A.T., Kleist O.A., Legina O.K., Belyakova N.V., Pantina R.A., Naryzhny S.N. (2018) In search of specific markers of glioblastoma: Analysis of proteoforms of glioblastoma cells. Tsitologiya, 60(7), 519-523. CrossRef Scholar google search
Naryzhny S., Ronzhina N., Zorina E., Kabachenko F., Klopov N., Zgoda V. (2022) Construction of 2DE patterns of plasma proteins: Aspect of potential tumor markers. Int. J. Mol. Sci., 23(19), 11113. CrossRef Scholar google search
Bradford M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72, 248-254. CrossRef Scholar google search
Naryzhny S.N., Zgoda V.G., Maynskova M.A., Novikova S.E., Ronzhina N.L., Vakhrushev I.V., Khryapova E.V., Lisitsa A.V., Tikhonova O.V., Ponomarenko E.A., Archakov A.I. (2016) Combination of virtual and experimental 2DE together with ESI LC-MS/MS gives a clearer view about proteomes of human cells and plasma. Electrophoresis, 37(2), 302-309. CrossRef Scholar google search
Naryzhny S.N. (1996) Upside-down stopped-flow electrofractionation of complex protein mixtures. Anal. Biochem., 238(1), 50-53. CrossRef Scholar google search
Naryzhny S.N. (2009) Blue Dry Western: Simple, economic, informative, and fast way of immunodetection. Anal. Biochem., 392(1), 90-95. CrossRef Scholar google search
Naryzhny S., Volnitskiy A., Kopylov A., Zorina E., Kamyshinsky R., Bairamukov V., Garaeva L., Shlikht A., Shtam T. (2020) Proteome of glioblastoma-derived exosomes as a source of biomarkers. Biomedicines, 8(7), 216. CrossRef Scholar google search
Naryzhny S.N., Legina O.K. (2022) Zonulin — regulation of tight contacts in the brain and intestine — facts and hypotheses. Biomeditsinskaya Khimiya, 68(5), 309-320. CrossRef Scholar google search
Hoagland IV L.F.M., Campa M.J., Gottlin E.B., Herndon J.E., Patz E.F. (2007) Haptoglobin and posttranslational glycan-modified derivatives as serum biomarkers for the diagnosis of nonsmall cell lung cancer. Cancer, 110(10), 2260-2268. CrossRef Scholar google search
Retrived February 25, 2024 from https://www.phosphosite.org/homeAction.action. Scholar google search
Aebersold R., Agar J.N., Amster I.J., Baker M.S., Bertozzi C.R., Boja E.S., Costello C.E., Cravatt B.F., Fenselau C., Garcia B.A., Ge Y., Gunawardena J., Hendrickson R.C., Hergenrother P.J., Huber C.G., Ivanov A.R., Jensen O.N., Jewett M.C., Kelleher N.L., Kiessling L.L., Krogan N.J., Larsen M.R., Loo J.A., Ogorzalek Loo R.R., Lundberg E., Maccoss M.J., Mallick P., Mootha V.K., Mrksich M., Muir T.W., Patrie S.M., Pesavento J.J., Pitteri S.J., Rodriguez H., Saghatelian A., Sandoval W., Schlüter H., Sechi S., Slavoff S.A., Smith L.M., Snyder M.P., Thomas P.M., Uhlén M., van Eyk J.E., Vidal M., Walt D.R., White F.M., Williams E.R., Wohlschlager T., Wysocki V.H., Yates N.A., Young N.L., Zhang B. (2018) How many human proteoforms are there? Nat. Chem. Biol., 14, 206-214. CrossRef Scholar google search
Murray-Zmijewski F., Slee E.A., Lu X. (2008) A complex barcode underlies the heterogeneous response of p53 to stress. Nat. Rev. Mol. Cell Biol., 9, 702-712. CrossRef Scholar google search
Naryzhny S.N., Legina O.K. (2019) Structural-functional diversity of p53 proteoforms. Biomeditsinskaya Khimiya, 65(4), 263-276. CrossRef Scholar google search
de Hart C.J., Chahal J.S., Flint S.J., Perlman D.H. (2014) Extensive post-translational modification of active and inactivated forms of endogenous p53. Mol. Cell. Proteomics, 13, 1-17. CrossRef Scholar google search
