Synthesis of affinity ligands and radioactive probes for isolation and study of myo-inositol 1,4,5-trisphosphate binding proteins

Journal Article · Tue May 29 00:00:00 EDT 1990 · Biochemistry; (United States)

DOI:https://doi.org/10.1021/bi00473a029· OSTI ID:5267354

Jina, A N; Ralph, J; Ballou, C E ^[1]

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1. Univ. of California, Berkeley (USA)

To synthesize an affinity matrix for isolation of D-myo-inositol 1,4,5-trisphosphate binding proteins, racemic 3-cyclohexene-1-carboxyaldehyde was oxidized and converted to a mixture of trans-3,4-dihydroxycyclohexane-1-carboxylic acid methyl ester isomers, which was phosphorylated and separated into ({plus minus})-(1R,3R,4R)- and ({plus minus})-(1R,3S,4S)-trans-3,4-bis((diphenoxyphosphoryl)oxy)cyclohexane-1-carboxylic acid methyl esters. Each of these racemic compounds was hydrogenolyzed and reacted with ethylenediamine to give a monoamide, N-(2-aminoethyl)-bis(phosphonyloxy)cyclohexane-1-carboxamide, that was coupled to cyanogen bromide activated Sepharose 4B to provide the desired affinity matrices. The intermediate trans-3,4-bis((diphenoxyphosphoryl)oxy)cyclohexane-1-carboxylic acid methyl ester was also reduced with lithium borotritide to give the (hydroxyl({sup 3}H)methyl)cyclohexane derivative, which was phosphorylated and hydrogenolyzed to yield trans-3,4-bis(phosphonyloxy)-1-((phosphonyloxy)({sup 3}H)methyl)cyclohexane, a radiolabeled analogue of inositol 1,4,5-trisphosphate. The carboxamide was also coupled to 4-azidosalicylic acid, and the product was iodinated to provide a {sup 125}I-radiolabeled photoactivatable cross-linking derivative of cyclohexanediol bisphosphate.

• OSTI ID: 5267354
• Journal Information: Biochemistry; (United States), Vol. 29:21; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CATALYST SUPPORTS
CHEMICAL PREPARATION
INOSITOL
MOLECULAR BIOLOGY
PHOSPHORIC ACID ESTERS
PROTEINS
FRACTIONATION
BIOCHEMICAL REACTION KINETICS
CYCLOHEXANE
IODINE 125
LIGANDS
LIQUID COLUMN CHROMATOGRAPHY
NMR SPECTRA
THIN-LAYER CHROMATOGRAPHY
TRITIUM COMPOUNDS
ALKANES
BETA DECAY RADIOISOTOPES
CARBOHYDRATES
CHROMATOGRAPHY
CYCLOALKANES
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
ESTERS
HYDROCARBONS
HYDROGEN COMPOUNDS
INOSITOLS
INTERMEDIATE MASS NUCLEI
INTERNAL CONVERSION RADIOISOTOPES
IODINE ISOTOPES
ISOTOPES
KINETICS
MONOSACCHARIDES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
RADIOISOTOPES
REACTION KINETICS
SACCHARIDES
SEPARATION PROCESSES
SPECTRA
SYNTHESIS
550201* - Biochemistry- Tracer Techniques