Design and synthesis of fluorogenic substrates that target protein phosphatases

Tetrahedron Letters

Volumn 45, Issue 4, 2004, Pages 707-710

  Zhu, Qing ^a   Huang, Xuan ^a   Chen, Grace Y J ^a   Yao, Shao Q ^a  

^a NATIONAL UNIVERSITY OF SINGAPORE   (Singapore)

Author keywords

Enzyme; Fluorogenic; High throughput; Phosphatase; Proteomics

Indexed keywords

ALKALINE PHOSPHATASE; PHOSPHATASE; PHOSPHOPROTEIN PHOSPHATASE; SERINE; THREONINE; TYROSINE PHOSPHATASE; UNCLASSIFIED DRUG;

AMINO ACID SEQUENCE; ARTICLE; CHEMICAL ANALYSIS; CHEMICAL STRUCTURE; ENZYME ANALYSIS; ENZYME SUBSTRATE; FLUORESCENCE ANALYSIS; MOLECULAR PROBE; PROTEIN TARGETING; SYNTHESIS;

EID: 0346157348     PISSN: 00404039     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.tetlet.2003.11.058     Document Type: Article

References (17)

1. Ahn N.G., Resing K.A. Nat. Biotechnol. 19:2001;317-318 Chen G.Y.J., Uttamchandani M., Lue Y.P.R., Lesaicherre M.L., Yao S.Q. Curr. Top. Med. Chem. 3:2003;705-724. and references cited therein.
2. Ahn N.G., Resing K.A. Nat. Biotechnol. 19:2001;317-318 Chen G.Y.J., Uttamchandani M., Lue Y.P.R., Lesaicherre M.L., Yao S.Q. Curr. Top. Med. Chem. 3:2003;705-724. and references cited therein.
3. Pawson T., Scott J.D. Science. 278:1997;2075-2080.
4. Walsh C. Enzymatic Reaction Mechanisms. 1979;Freeman, New York.
5. Zhu Q., Huang X., Chen G.Y.J., Yao S.Q. Tetrahedron Lett. 44:2003;2669-2672 Wahler D., Badalassi F., Reymond J.L. Chem. Eur. J. 8:2002;3211-3228.
6. Zhu Q., Huang X., Chen G.Y.J., Yao S.Q. Tetrahedron Lett. 44:2003;2669-2672 Wahler D., Badalassi F., Reymond J.L. Chem. Eur. J. 8:2002;3211-3228.
7. Chen G.Y.J., Uttamchandani M., Zhu Q., Wang G., Yao S.Q. ChemBiochem. 4:2003;336-339 Zhu Q., Uttamchandani M., Li D.B., Lesaicherre M.L., Yao S.Q. Org. Lett. 5:2003;1257-1260.
8. Chen G.Y.J., Uttamchandani M., Zhu Q., Wang G., Yao S.Q. ChemBiochem. 4:2003;336-339 Zhu Q., Uttamchandani M., Li D.B., Lesaicherre M.L., Yao S.Q. Org. Lett. 5:2003;1257-1260.
9. Probe 1 to some extent is structurally similar to the two fluorogenic substrates sold by Molecular Probes (Eugene, USA): MUP (4-methylumbelliferone phosphate) and DiFMUP (6,8-difluoro-4-methylumbelliferone phosphate).
10. Coincidently, Probe 2 is structurally similar to the fluorogenic substrate published by Wahler et al., which was shown to be a substrate for alkaline phosphatases. See Refs. [4b,8] for more details.
11. Wahler D., Badalassi F., Crotti P., Reymond J.L. Angew. Chem., Int. Ed. 40:2001;4457-4460.
12. Various conditions (e.g., p-toluenesulfonyl chloride/DCM/DIEA or pyridine, methanesulfonyl chloride/DCM/DIEA or pyridine) were attempted. Only trace amounts of the desired product were detected (by TLC and MS). Mostly starting compound, 4 , was recovered.
13. + (100).
14. + (100).
15. A : Acid phosphatase from wheat germ (P-3627); B : acid phosphatase from potato (P-3752); C : alkaline phosphatase from bovine intestinal mucosa (P-7640); D : alkaline phosphatase from rabbit intestine (P-2265); E : alkaline phosphatase from porcine intestinal mucosa (P-4002); F : alkaline phosphate from shrimp (M182A 11302611); G : alkaline phosphatase from calf intestinal (M820A 13595209); H : PP1 Ser/Thr phosphatase (P0754S); I : LAR Tyr phosphatase (P0750S); J : Lambda PPase (P0753S); K : PTP (P0752S); L : YOP (P0751S).
16. 4 and BSA were tested.
17. The fluorogenic reaction was carried in a black round-bottom polypropylene 96-well microtitre plates (Nunc, USA). The fluorescence was detected using the SpectraMAX™ Gemini XS fluorescence plate reader (Molecular Devices, USA).