Identification of Peptide Substrate and Small Molecule Inhibitors of Testis-Specific Serine/Threonine Kinase1 (TSSK1) By the Developed Assays

Zhang, Leilei; Yu, Yongping; Liu, Zijie; Abliz, Zeper; Liu, Gang

doi:10.1021/jm9002846

Cited by 11 publications

(10 citation statements)

References 40 publications

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“…On the other hand, TSSK1 has been shown to phosphorylate the AMARA peptide (AMARAASAAALARRR), a commonly used AMPK substrate [ 54 , 51 ], and it is predicted that all TSSKs will phosphorylate this substrate. Two peptides, Pep4 (PLSRTLSVSS) and Pep8 (AALVRQMSVAFFFK), have also been reported as suitable TSSK1 substrates for high-throughput screening (HTS) by luminescence detection [ 55 ]. TSSK6 is able to phosphorylate the GKGRGLSLARFAKK peptide from the myelin basic protein (MBP) sequence [ 52 ].…”

Section: In Vitro Biochemical Characterization Of Tssk Kinasesmentioning

confidence: 99%

Testis-specific serine kinase protein family in male fertility and as targets for non-hormonal male contraception†

Salicioni

Gervasi

Sosnik

et al. 2020

Biology of Reproduction

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Male contraception is a very active area of research. Several hormonal agents have entered clinical trials, while potential non-hormonal targets have been brought to light more recently and are at earlier stages of development. The general strategy is to target genes along the molecular pathways of sperm production, maturation, or function, and it is predicted that these novel approaches will hopefully lead to more selective male contraceptive compounds with a decreased side effect burden. Protein kinases are known to play a major role in signaling events associated with sperm differentiation and function. In this review, we focus our analysis on the testis-specific serine kinase (TSSK) protein family. We have previously shown that members of the family of TSSKs are postmeiotically expressed in male germ cells and in mature mammalian sperm. The restricted postmeiotic expression of TSSKs as well as the importance of phosphorylation in signaling processes strongly suggests that TSSKs have an important role in germ cell differentiation and/or sperm function. This prediction has been supported by the reported sterile phenotype of the Tssk6 knockout (KO) mice and of the double Tssk1 and Tssk2 KO mice and by the male subfertile phenotype observed in a Tssk4 KO mouse model. Summary Sentence The established role of TSSKs in spermiogenesis and in male fertility, together with their unique kinase features, strongly supports this family of protein kinases as a very promising drug discovery target for development of a non-hormonal male contraceptive.

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Section: In Vitro Biochemical Characterization Of Tssk Kinasesmentioning

confidence: 99%

Testis-specific serine kinase protein family in male fertility and as targets for non-hormonal male contraception†

Salicioni

Gervasi

Sosnik

et al. 2020

Biology of Reproduction

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“…In order to determine the compounds by ESI‐MS without performing sample pretreatment, an online desalting system was adopted in this study. A switching valve on the Waters UPLC/TQMS system was successfully combined with efficient LC separation to completely remove non‐volatile additives, such as Tris‐HCl, DTT, and MgCl 2 , from the kinase reaction mixtures prior to analysis by MS . The early eluent (before 1.2 min, Fig.…”

Section: Resultsmentioning

confidence: 99%

“…During this process, the synthetic peptide PKTPKKAKKL (an excellent substrate for cdk5/p25) was applied in the enzymatic reaction. Compared with other existing analytical methods, the UPLC/TQMS method can detect the products or substrates directly and quantitatively, avoiding modification of substrates or secondary enzymatic reactions that are irrelevant to target enzyme reaction …”

mentioning

confidence: 99%

Determination of kinetic parameters and structure–activity relationships of ginsenosides as inhibitors of cyclin‐dependent kinase 5/p25 using ultra‐pressure liquid chromatography with triple quadrupole tandem mass spectrometry

Chen

Yang

Niu

et al. 2013

Rapid Comm Mass Spectrometry

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“…Compound 2 is reported on the PubChem website (pubchem.ncbi.nlm.nih.gov) to have demonstrable activity in 13 bioassays, including an additional kinase assay, glycogen synthase kinase 3 beta, as well as being cited in the literature as inhibiting testis-specific serine/ threonine kinase 1. 15 This suggests that compound 2 may be binding a structural motif common to serine/threonine kinases. Although compound 3 has no previously reported biological activity and low-micromolar in vitro potency, in our cell-based assay for p38 inhibition, it appears to reduce the phosphorylation of ERK1/2 in addition to MNK1, indicating that it does not have specificity for p38 (data not shown).…”

Section: Discussionmentioning

confidence: 99%

“…[15][16][17] Such modifications could result in a better correspondence between biochemical and cellular potency. Furthermore, the measured ITC data indicate that there is a discrete unimolecular binding site for compound 1, providing further support for crystallography studies, rational drug design, and structural-activity relationship studies.…”

Section: Discussionmentioning

confidence: 99%

Discovery and Characterization of a Biologically Active Non–ATP-Competitive p38 MAP Kinase Inhibitor

et al. 2016

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Mitogen-activated protein kinase (MAPK) p38 is part of a broad and ubiquitously expressed family of MAPKs whose activity is responsible for mediating an intracellular response to extracellular stimuli through a phosphorylation cascade. p38 is central to this signaling node and is activated by upstream kinases while being responsible for activating downstream kinases and transcription factors via phosphorylation. Dysregulated p38 activity is associated with numerous autoimmune disorders and has been implicated in the progression of several types of cancer. A number of p38 inhibitors have been tested in clinical trials, with none receiving regulatory approval. One characteristic shared by all of the compounds that failed clinical trials is that they are all adenosine triphosphate (ATP)-competitive p38 inhibitors. Seeing this lack of mechanistic diversity as an opportunity, we screened ~32,000 substances in search of novel p38 inhibitors. Among the inhibitors discovered is a compound that is both non-ATP competitive and biologically active in cell-based models for p38 activity. This is the first reported discovery of a non-ATP-competitive p38 inhibitor that is active in cells and, as such, may enable new pharmacophore designs for both therapeutic and basic research to better understand and exploit non-ATPcompetitive inhibitors of p38 activity.

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Identification of Peptide Substrate and Small Molecule Inhibitors of Testis-Specific Serine/Threonine Kinase1 (TSSK1) By the Developed Assays

Cited by 11 publications

References 40 publications

Testis-specific serine kinase protein family in male fertility and as targets for non-hormonal male contraception†

Testis-specific serine kinase protein family in male fertility and as targets for non-hormonal male contraception†

Determination of kinetic parameters and structure–activity relationships of ginsenosides as inhibitors of cyclin‐dependent kinase 5/p25 using ultra‐pressure liquid chromatography with triple quadrupole tandem mass spectrometry

Discovery and Characterization of a Biologically Active Non–ATP-Competitive p38 MAP Kinase Inhibitor

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