Vol.:(0123456789) 1 3 Molecular and Cellular Biochemistry https://doi.org/10.1007/s11010-020-03747-1 CIGB‑300 anticancer peptide regulates the protein kinase CK2‑dependent phosphoproteome Yasser Perera 1,2  · Yassel Ramos 3  · Gabriel Padrón 3  · Evelin Caballero 2  · Osmany Guirola 3  · Lorena G. Caligiuri 4  · Norailys Lorenzo 4  · Florencia Gottardo 4  · Hernán G. Farina 4  · Odile Filhol 5  · Claude Cochet 5  · Silvio E. Perea 2 Received: 13 January 2020 / Accepted: 6 May 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020 Abstract Casein-kinase CK2 is a Ser/Thr protein kinase that fosters cell survival and proliferation of malignant cells. The CK2 holo- enzyme, formed by the association of two catalytic alpha/alpha’ (CK2α/CK2α’) and two regulatory beta subunits (CK2β), phosphorylates diverse intracellular proteins partaking in key cellular processes. A handful of such CK2 substrates have been identifed as targets for the substrate-binding anticancer peptide CIGB-300. However, since CK2β also contains a CK2 phosphorylation consensus motif, this peptide may also directly impinge on CK2 enzymatic activity, thus globally modifying the CK2-dependent phosphoproteome. To address such a possibility, frstly, we evaluated the potential interaction of CIGB- 300 with CK2 subunits, both in cell-free assays and cellular lysates, as well as its efect on CK2 enzymatic activity. Then, we performed a phosphoproteomic survey focusing on early inhibitory events triggered by CIGB-300 and identifed those CK2 substrates signifcantly inhibited along with disturbed cellular processes. Altogether, we provided here the frst evidence for a direct impairment of CK2 enzymatic activity by CIGB-300. Of note, both CK2-mediated inhibitory mechanisms of this anticancer peptide (i.e., substrate- and enzyme-binding mechanism) may run in parallel in tumor cells and help to explain the diferent anti-neoplastic efects exerted by CIGB-300 in preclinical cancer models. Keywords CIGB-300 · CK2 · Anticancer peptides · CPP · Clinical-grade CK2 inhibitor · Phosphoproteomics Introduction Casein-kinase 2 (CK2) seems to account for about 20% of the cellular phosphoproteome in living cells [1]. This enzyme is usually deregulated in malignant cells and has been linked to hallmarks of cancer including exacerbated cell proliferation, increased survival, angiogenesis, and metastasis [2]. Of note, CK2 modulates the activity of substrates with driver roles in cancer including both tumor suppressors like PTEN [3] and PML [4], as well as oncogenes like AKT [5] and c-myc [6]. Moreover, aberrant CK2 expression impacts on several signaling pathways whose deregulation leads to malignant transformation including Wnt signaling, Hedgehog signaling (Hh), JAK/STAT, and PI3K/AKT pathway [7]. CK2 enzymatic activity is exerted by the catalytic subu- nits alone and/or the full holoenzyme which is composed of two catalytic CK2α/α’ and two regulatory CK2β subunits [8]. Phosphorylation of a subset of substrates can only be exerted by the holoenzyme CK2α 2 β 2 (class-III substrates), whereas class-I substrates can be phosphorylated either by CK2α 2 β 2 or the free catalytic subunits. A third class of CK2 Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11010-020-03747-1) contains supplementary material, which is available to authorized users. * Yasser Perera yasserperera@outlook.com 1 China-Cuba Biotechnology Joint Innovation Center (CCBJIC), Yongzhou Zhong Gu Biotechnology Co., Ltd, Yangjiaqiao Street, Lengshuitan District, Yongzhou City 425000, Hunan Province, China 2 Molecular Oncology Group, Division of Pharmaceuticals, Center for Genetic Engineering & Biotechnology, Ave 31 e/158 & 190, Playa, 10600 Havana, Cuba 3 Department of Proteomics. Biomedical Research Division, Center for Genetic Engineering & Biotechnology, Havana, Cuba 4 Platform of Biotechnology Services, Quilmes National University, Roque Saenz Peña 352, 1876 Bernal, Buenos Aires, Argentina 5 University of Grenoble Alpes, Inserm U1036, CEA, IRIG-BCI, 38000 Grenoble, France