Photoaffinity Heterobifunctional Cross-Linking Reagents Based on Azide-Substituted Salicylates

Nobuyuki Imai; Lori D. Dwyer; Tadashi Kometani; Tae Ji; Thomas C. Vanaman; David S. Watt

doi:10.1021/bc00002a009

Photoaffinity Heterobifunctional Cross-Linking Reagents Based on Azide-Substituted Salicylates

Nobuyuki Imai, Lori D. Dwyer, Tadashi Kometani, Tae Ji, Thomas C. Vanaman, David S. Watt

Molecular and Cellular Biochemistry

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

4 Scopus citations

Abstract

Photoaffinity heterobifunctional cross-linking reagents are described that incorporate a 4-azidosali-cylate group at one terminus and an N-hydroxysuccinimidyl ester at the other terminus with a “linking arm” of variable length separating the photoactive and electrophilic termini. Exposure of calmodulin (CaM) to succinimidyl N-[2-[(4-azidosalicyloyl)oxy]ethyl]suberamate (4C) led to monoad-ducts at Lys-21, Lys-75, and Lys-94. Separation of the monoadducts from CaM and polyadducts, radioiodination, and photolysis in the presence of human erythrocyte plasma membrane Ca²⁺, Mg²⁺-ATPase led to calcium-dependent cross-linking with 8% cross-linking efficiency.

Original language	English
Pages (from-to)	144-148
Number of pages	5
Journal	Bioconjugate Chemistry
Volume	1
Issue number	2
DOIs	https://doi.org/10.1021/bc00002a009
State	Published - Mar 1 1990

Bibliographical note

Funding Information:
We thank the National Science Foundation (Grant CHE- 8607441) for financial support, Degussa AG for a generous gift of amino acids, and the University of Kentucky for the purchase of bond issue equipment.

Funding Information:
ACKNOWLEDGMENT We thank the National Science Foundation (Grant CHE- 8607441) for financial support, Degussa AG for a gener- ousgiftofaminoacids,andtheUniversityofKentucky forthepurchaseofbondissueequipment.

ASJC Scopus subject areas

Biotechnology
Bioengineering
Biomedical Engineering
Pharmacology
Pharmaceutical Science
Organic Chemistry

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10.1021/bc00002a009

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title = "Photoaffinity Heterobifunctional Cross-Linking Reagents Based on Azide-Substituted Salicylates",

abstract = "Photoaffinity heterobifunctional cross-linking reagents are described that incorporate a 4-azidosali-cylate group at one terminus and an N-hydroxysuccinimidyl ester at the other terminus with a “linking arm” of variable length separating the photoactive and electrophilic termini. Exposure of calmodulin (CaM) to succinimidyl N-[2-[(4-azidosalicyloyl)oxy]ethyl]suberamate (4C) led to monoad-ducts at Lys-21, Lys-75, and Lys-94. Separation of the monoadducts from CaM and polyadducts, radioiodination, and photolysis in the presence of human erythrocyte plasma membrane Ca2+, Mg2+-ATPase led to calcium-dependent cross-linking with 8% cross-linking efficiency.",

author = "Nobuyuki Imai and Dwyer, {Lori D.} and Tadashi Kometani and Tae Ji and Vanaman, {Thomas C.} and Watt, {David S.}",

note = "Funding Information: We thank the National Science Foundation (Grant CHE- 8607441) for financial support, Degussa AG for a generous gift of amino acids, and the University of Kentucky for the purchase of bond issue equipment. Funding Information: ACKNOWLEDGMENT We thank the National Science Foundation (Grant CHE- 8607441) for financial support, Degussa AG for a gener- ousgiftofaminoacids,andtheUniversityofKentucky forthepurchaseofbondissueequipment.",

year = "1990",

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doi = "10.1021/bc00002a009",

language = "English",

volume = "1",

pages = "144--148",

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T1 - Photoaffinity Heterobifunctional Cross-Linking Reagents Based on Azide-Substituted Salicylates

AU - Imai, Nobuyuki

AU - Dwyer, Lori D.

AU - Kometani, Tadashi

AU - Ji, Tae

AU - Vanaman, Thomas C.

AU - Watt, David S.

N1 - Funding Information: We thank the National Science Foundation (Grant CHE- 8607441) for financial support, Degussa AG for a generous gift of amino acids, and the University of Kentucky for the purchase of bond issue equipment. Funding Information: ACKNOWLEDGMENT We thank the National Science Foundation (Grant CHE- 8607441) for financial support, Degussa AG for a gener- ousgiftofaminoacids,andtheUniversityofKentucky forthepurchaseofbondissueequipment.

PY - 1990/3/1

Y1 - 1990/3/1

N2 - Photoaffinity heterobifunctional cross-linking reagents are described that incorporate a 4-azidosali-cylate group at one terminus and an N-hydroxysuccinimidyl ester at the other terminus with a “linking arm” of variable length separating the photoactive and electrophilic termini. Exposure of calmodulin (CaM) to succinimidyl N-[2-[(4-azidosalicyloyl)oxy]ethyl]suberamate (4C) led to monoad-ducts at Lys-21, Lys-75, and Lys-94. Separation of the monoadducts from CaM and polyadducts, radioiodination, and photolysis in the presence of human erythrocyte plasma membrane Ca2+, Mg2+-ATPase led to calcium-dependent cross-linking with 8% cross-linking efficiency.

AB - Photoaffinity heterobifunctional cross-linking reagents are described that incorporate a 4-azidosali-cylate group at one terminus and an N-hydroxysuccinimidyl ester at the other terminus with a “linking arm” of variable length separating the photoactive and electrophilic termini. Exposure of calmodulin (CaM) to succinimidyl N-[2-[(4-azidosalicyloyl)oxy]ethyl]suberamate (4C) led to monoad-ducts at Lys-21, Lys-75, and Lys-94. Separation of the monoadducts from CaM and polyadducts, radioiodination, and photolysis in the presence of human erythrocyte plasma membrane Ca2+, Mg2+-ATPase led to calcium-dependent cross-linking with 8% cross-linking efficiency.

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Photoaffinity Heterobifunctional Cross-Linking Reagents Based on Azide-Substituted Salicylates

Abstract

Bibliographical note

ASJC Scopus subject areas

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