The protonated form of 1-N6-etheno-[erythro-9-(2-hydroxy-3-nonyl)] adenine is identified at the active site of adenosine deaminase

Valeria R. Caiolfa; David Gill; Abraham H. Parola

doi:10.1016/0014-5793(90)80055-N

The protonated form of 1-N⁶-etheno-[erythro-9-(2-hydroxy-3-nonyl)] adenine is identified at the active site of adenosine deaminase

Valeria R. Caiolfa, David Gill, Abraham H. Parola

Department of Chemistry

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

A novel fluorescent competitive inhibitor of adenosine deaminase (EC 3.5.4.4) (ADA), 1-N⁶-etheno-[erythro-9-(2-hydroxy-3-nonyl)] adenine (ε{lunate}-EHNA), is protonated at the active site of the enzyme. In ε{lunate}-EHNA [K₁ = (4.06 ± 1.00) 10^-6 M] part of the competive inhibition of EHNA is combined with spectroscopic properties of etheno-adenines. Computer subtraction of the fluorescence excitation spectrum of ADA from that of its equimolar complex with ε{lunate}-EHNA yielded the corrected excitation spectrum of ε{lunate}-EHNA at the active site of the enzyme. This spectrum mimics that of ε{lunate}-EHNA at pH 5.5 in buffer solution and is suggested to indicate a shift in protonation equilibrium at the active site.

Original language	English
Pages (from-to)	19-22
Number of pages	4
Journal	FEBS Letters
Volume	260
Issue number	1
DOIs	https://doi.org/10.1016/0014-5793(90)80055-N
State	Published - 15 Jan 1990

Keywords

1-N-Etheno-[erythro-9-(2-hydroxy-3-nonyl)] adenine
Adenosine deaminase
Etheno-adenine
Fluorescence polarization
Fluorescent inhibitor
pH indicator

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

Access to Document

10.1016/0014-5793(90)80055-N

Cite this

@article{cc702a49fcd04f9d88a7fe9453f86957,

title = "The protonated form of 1-N6-etheno-[erythro-9-(2-hydroxy-3-nonyl)] adenine is identified at the active site of adenosine deaminase",

abstract = "A novel fluorescent competitive inhibitor of adenosine deaminase (EC 3.5.4.4) (ADA), 1-N6-etheno-[erythro-9-(2-hydroxy-3-nonyl)] adenine (ε{lunate}-EHNA), is protonated at the active site of the enzyme. In ε{lunate}-EHNA [K1 = (4.06 ± 1.00) 10-6 M] part of the competive inhibition of EHNA is combined with spectroscopic properties of etheno-adenines. Computer subtraction of the fluorescence excitation spectrum of ADA from that of its equimolar complex with ε{lunate}-EHNA yielded the corrected excitation spectrum of ε{lunate}-EHNA at the active site of the enzyme. This spectrum mimics that of ε{lunate}-EHNA at pH 5.5 in buffer solution and is suggested to indicate a shift in protonation equilibrium at the active site.",

keywords = "1-N-Etheno-[erythro-9-(2-hydroxy-3-nonyl)] adenine, Adenosine deaminase, Etheno-adenine, Fluorescence polarization, Fluorescent inhibitor, pH indicator",

author = "Caiolfa, {Valeria R.} and David Gill and Parola, {Abraham H.}",

note = "Funding Information: Ack~owie~ge~e~Wts:e are indebtedt o Mr Max Enkin for having writtent he computerp rograma nd to ProfessorsS . Bitner, D. Kost and J. Beckerf or their advicea nd help. This work was supportedin part by grantN OOO14-89-J-16o2f5 t he Office of Naval Research.",

year = "1990",

month = jan,

day = "15",

doi = "10.1016/0014-5793(90)80055-N",

language = "English",

volume = "260",

pages = "19--22",

journal = "FEBS Letters",

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publisher = "Wiley-Blackwell",

number = "1",

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T1 - The protonated form of 1-N6-etheno-[erythro-9-(2-hydroxy-3-nonyl)] adenine is identified at the active site of adenosine deaminase

AU - Caiolfa, Valeria R.

AU - Gill, David

AU - Parola, Abraham H.

N1 - Funding Information: Ack~owie~ge~e~Wts:e are indebtedt o Mr Max Enkin for having writtent he computerp rograma nd to ProfessorsS . Bitner, D. Kost and J. Beckerf or their advicea nd help. This work was supportedin part by grantN OOO14-89-J-16o2f5 t he Office of Naval Research.

PY - 1990/1/15

Y1 - 1990/1/15

N2 - A novel fluorescent competitive inhibitor of adenosine deaminase (EC 3.5.4.4) (ADA), 1-N6-etheno-[erythro-9-(2-hydroxy-3-nonyl)] adenine (ε{lunate}-EHNA), is protonated at the active site of the enzyme. In ε{lunate}-EHNA [K1 = (4.06 ± 1.00) 10-6 M] part of the competive inhibition of EHNA is combined with spectroscopic properties of etheno-adenines. Computer subtraction of the fluorescence excitation spectrum of ADA from that of its equimolar complex with ε{lunate}-EHNA yielded the corrected excitation spectrum of ε{lunate}-EHNA at the active site of the enzyme. This spectrum mimics that of ε{lunate}-EHNA at pH 5.5 in buffer solution and is suggested to indicate a shift in protonation equilibrium at the active site.

AB - A novel fluorescent competitive inhibitor of adenosine deaminase (EC 3.5.4.4) (ADA), 1-N6-etheno-[erythro-9-(2-hydroxy-3-nonyl)] adenine (ε{lunate}-EHNA), is protonated at the active site of the enzyme. In ε{lunate}-EHNA [K1 = (4.06 ± 1.00) 10-6 M] part of the competive inhibition of EHNA is combined with spectroscopic properties of etheno-adenines. Computer subtraction of the fluorescence excitation spectrum of ADA from that of its equimolar complex with ε{lunate}-EHNA yielded the corrected excitation spectrum of ε{lunate}-EHNA at the active site of the enzyme. This spectrum mimics that of ε{lunate}-EHNA at pH 5.5 in buffer solution and is suggested to indicate a shift in protonation equilibrium at the active site.

KW - 1-N-Etheno-[erythro-9-(2-hydroxy-3-nonyl)] adenine

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