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Fluorescence detection techniques for protein kinase assay

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Abstract

Traditional methods for protein kinase (PK) assay are mainly based on use of 32P-labeled adenosine triphosphate (ATP); applications of such methods are, however, hampered by radioactive waste and short half-life of 32P-labeled ATP. Therefore non-radioactive methods, such as fluorescence detection techniques are good alternative. In this review, we describe the principles of four fluorescence techniques (fluorescence intensity endpoint measurement, fluorescence resonance energy transfer (FRET), fluorescence polarization (FP), and fluorescence lifetime imaging) and provide an overview of applications of these fluorescence detection techniques in protein kinase assay, underlining their relative advantages and limitations. Research trends in this field are also highlighted.

Schematic representation of kinase assay based on direct fluorescence polarization measurements. The fluorescent peptide, on phosphorylation by kinase, binds to a phosphospecific antibody, which leads to a high FP value

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Abbreviations

PK:

Protein kinase

ATP:

Adenosine triphosphate

FRET:

Fluorescence resonance energy transfer

FP:

Fluorescence polarization

NMR:

Nuclear magnetic resonance

MS:

Mass spectrometry

RP-HPLC:

Reversed-phase high-performance liquid chromatography

CZE:

Capillary zone electrophoresis

GFP:

Green fluorescence protein

CFP:

Cyan fluorescent protein

YFP:

Yellow fluorescent protein

APC:

Algae protein allophycocyanin

FLIM:

Fluorescence lifetime imaging microscope

EGFR:

Epidermal growth factor receptor

PDGF:

Platelet-derived growth factor

LRP:

Lipoprotein receptor-related protein

References

  1. Sahal D, Fujita-Yamaguchi Y (1987) Anal Biochem 167:23–30

    Article  CAS  Google Scholar 

  2. Glass DB, Masaracchia RA, Feramisco JR, Kemp BE (1978) Anal Biochem 87:566–575

    Article  CAS  Google Scholar 

  3. Witt JJ, Roskoski R (1975) Anal Biochem 66:253–258

    Article  CAS  Google Scholar 

  4. Keane NE, Chavanieu A, Quirk PG, Evans JS, Levine BA, Calas B, Wei L, Ellis L (1994) Eur J Biochem 226:525–536

    Article  CAS  Google Scholar 

  5. Newton RP, Evans AM, Langridge JI, Walton TJ, Harris FM, Brenton AG (1995) Anal Biochem 224:32–38

    Article  CAS  Google Scholar 

  6. Cann AD, Wolf I, Kohanski RA (1997) Anal Biochem 247:327–332

    Article  CAS  Google Scholar 

  7. Dawson JF, Boland MP, Holmes CFB (1994) Anal Biochem 220:340–345

    Article  CAS  Google Scholar 

  8. Wang ZX, Cheng Q, Killilea SD (1995) Anal Biochem 230:55–61

    Article  CAS  Google Scholar 

  9. Barker SC, Kassel DB, Weigl D, Huang X, Luther MA, Knight WB (1995) Biochemistry 34:14843–14851

    Article  CAS  Google Scholar 

  10. Koyama-Honda I, Ritchie K, Fujiwara T, Iino R, Murakoshi H, Kasai RS, Kusumi A (2005) Biophys J 88:2126–2136

    Article  CAS  Google Scholar 

  11. Day RN, Schaufele F (2005) Mol Endocrinol 19:1675–1686

    Article  CAS  Google Scholar 

  12. Karvinen J, Laitala V, Makinen ML, Mulari O, Tamminen J, Hermonen J, Hurskainen P, Hemmila I (2004) Anal Chem 76:1429–1436

    Article  CAS  Google Scholar 

  13. Yan Y, Marriott G (2003) Curr Opin Chem Biol 7:635–640

    Article  CAS  Google Scholar 

  14. Perrin F (1926) J Phys Radium 7:390–401

    Article  CAS  Google Scholar 

  15. Eremin SA, Gallacher G, Lotey H, Smith DS, Landon J (1987) Clin Chem 33:1903–1906

    CAS  Google Scholar 

  16. Aucouturier P, Preud’homme JL, Lubochinsky B (1983) Diagn Immunol 1:310–314

    CAS  Google Scholar 

  17. Murakami A, Nakaura M, Nakatsuji Y, Nagahara S, Tran-Cong Q, Makino K (1991) Nucleic Acids Res 19:4097–4102

    Article  CAS  Google Scholar 

  18. Kinoshita K, Maeda H, Hinuma Y (1980) Anal Biochem 104:15–22

    Article  CAS  Google Scholar 

  19. Seethala R, Menzel R (1997) Anal Biochem 253:210–218

    Article  CAS  Google Scholar 

  20. Kristjansdottir K, Rudolph J (2003) Anal Biochem 316:41–49

    Article  CAS  Google Scholar 

  21. Seethala R, Menzel R (1998) Anal Biochem 255:257–262

    Article  CAS  Google Scholar 

  22. Turek TC, Small EC, Bryant RW, Adam W, Hill G (2001) Anal Biochem 299:45–53

    Article  CAS  Google Scholar 

  23. Coffin J, Latev M, Bi X, Nikiforov TT (2000) Anal Biochem 278:206–212

    Article  CAS  Google Scholar 

  24. Turek-Etienne TC, Kober TP, Stafford JM, Bryant RW (2003) Assay Drug Dev Technol 1:545–553

    Article  CAS  Google Scholar 

  25. Scott JE, Carpenter JW (2003) Anal Biochem 316:82–91

    Article  CAS  Google Scholar 

  26. Clegg RM (1992) Methods Enzymol 211:353–388

    Article  CAS  Google Scholar 

  27. Sato M, Umezawa Y (2004) Methods 32:451–455

    Article  CAS  Google Scholar 

  28. Mathis G (1995) Clin Chem 41:1391–1397

    CAS  Google Scholar 

  29. Li Y, Cummings RT, Cunningham BR, Chen Y, Zhou G (2003) Anal Biochem 321:151–156

    Article  CAS  Google Scholar 

  30. Zhang WX, Wang R, Wisniewski D, Marcy AI, LoGrasso P, Lisnock J-M, Cummings RT, Thompson JE (2005) Anal Biochem 343:76–83

    CAS  Google Scholar 

  31. Riddle SM, Vedvik KL, Hanson GT, Vogel KW (2006) Anal Biochem 356:108–116

    Article  CAS  Google Scholar 

  32. Rodems SM, Hamman BD, Lin C, Zhao J, Shah S, Heidary D, Makings L, Stack JH, Pollok BA (2002) Assay Drug Dev Technol 1:9–19

    Article  CAS  Google Scholar 

  33. Jia Y, Quinn CM, Gagnon AI, Talanian R (2006) Anal Biochem 356:273–281

    Article  CAS  Google Scholar 

  34. McIlroy BK, Walters JD, Johnson JD (1991) Anal Biochem 195:148–152

    Article  CAS  Google Scholar 

  35. Lutz MP, Pinon DI, Miller LJ (1994) Anal Biochem 220:268–274

    Article  CAS  Google Scholar 

  36. Isbell JC, Christian ST, Mashburn NA, Bell PD (1995) Life Sci 57:1701–1707

    Article  CAS  Google Scholar 

  37. Sano M, Ueno K, Kamimori H (2003) J Chromatogr B 794:149–156

    Article  CAS  Google Scholar 

  38. Zarrine-Afsar A, Krylov SN (2003) Anal Chem 75:3720–3724

    Article  CAS  Google Scholar 

  39. Kupcho K, Somberg R, Bulleit B, Goueli SA (2003) Anal Biochem 317:210–217

    Article  CAS  Google Scholar 

  40. Rininsland F, Xia W, Wittenburg S, Shi X, Stankewicz C, Achyuthan K, McBranch D, Whitten D (2004) Proc Natl Acad Sci USA 101:15295–15300

    Article  CAS  Google Scholar 

  41. Andersson L, Porath L (1986) Anal Biochem 154:250–254

    Article  CAS  Google Scholar 

  42. Houseman BT, Huh JH, Kron SJ, Mrksich M (2002) Nat Biotechnol 20:270–274

    Article  CAS  Google Scholar 

  43. Lesaicherre ML, Uttamchandani M, Chen GY, Yao SQ (2002) Bioorg Med Chem Lett 12:2085–2088

    Article  CAS  Google Scholar 

  44. Martin K, Steinberg TH, Cooley LA, Gee KR, Beechem JM, Patton WF (2003) Proteomics 3:1244–1255

    Article  CAS  Google Scholar 

  45. Lakowicz JR, Szmacinski H, Nowaczyk K, Lederer WJ, Kirby MS, Johnson ML (1994) Cell Calcium 15:7–27

    Article  CAS  Google Scholar 

  46. Mahajan NP, Linder K, Berry G, Gordon GW, Heim R, Herman B (1998) Nat Biotechnol 16:547–552

    Article  CAS  Google Scholar 

  47. Miyawaki A, Llopis J, Heim R, McCaffery JM, Adams JA, Ikura M, Tsien RY (1997) Nature 388:882–887

    Article  CAS  Google Scholar 

  48. Duncan RR (2006) Biochem Soc Trans 34:679–682

    Article  CAS  Google Scholar 

  49. Wouters FS, Bastiaens PI (1999) Curr Biol 9:1127–1130

    Article  CAS  Google Scholar 

  50. Calleja V, Ameer-Beg S, Vojnovic B, Woscholski R, Downward J, Larijani B (2003) Biochem J 372:33–40

    Article  CAS  Google Scholar 

  51. Peltan ID, Thomas AV, Mikhailenko I, Strickland DK, Hyman BT, Arnim CAFV (2006) Biochem Biophys Res Commun 349:24–30

    Article  CAS  Google Scholar 

  52. Lebakken CS, Kang HC, Vogel KW (2007) J Biomol Screen 12:828–841

    Article  CAS  Google Scholar 

  53. Inamori K, Kyo M, Nishiya Y, Inoue Y, Sonoda T, Kinoshita E, Koike T, Katayama Y (2005) Anal Chem 77:3979–3985

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. College of Bioengineering, Hubei University of Technology, Wuhan, 430068, China

    YongJin Li, WeiHong Xie & GuiJie Fang

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  1. YongJin Li
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  2. WeiHong Xie
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  3. GuiJie Fang
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Correspondence to YongJin Li.

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Li, Y., Xie, W. & Fang, G. Fluorescence detection techniques for protein kinase assay. Anal Bioanal Chem 390, 2049–2057 (2008). https://doi.org/10.1007/s00216-008-1986-z

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