公开(公告)号：WO1996036729A1

公开(公告)日：1996-11-21

申请号：PCT/US1996006860

申请日：1996-05-14

Applicant: COULTER INTERNATIONAL CORP.

Inventor： COULTER INTERNATIONAL CORP. ,  LUCAS, Frank, J. ,  JAFFE, Gerald, E. ,  BOTT, Steven, E. ,  CARTER, James, H.

IPC: C12Q01/34

CPC classification number: C12Q1/34 ,  C12Q2334/40 ,  C12Q2337/40 ,  G01N33/574

Abstract: An assay compound or a salt thereof for assaying the activity of an enzyme inside a metabolically active whole cell is disclosed. The assay compound includes a leaving group and an indicator group. The leaving group is selected from the group comprising amino acids, peptides, saccharides, sulfates, phosphates, esters, phosphate esters, nucleotides, polynucleotides, nucleic acids, pyrimidines, purines, nucleosides, lipids and mixtures thereof. The indicator group is selected from compounds which have a first state when joined to the leaving group, and a second state when the leaving group is cleaved from the indicator group by the enzyme. Preferably, the indicator compounds are rhodamine 110, rhodol, and fluorescein and analogs of these compounds. A method of synthesizing the compound as well as methods of using these compounds to measure enzyme activity are also disclosed.

Abstract translation: 公开了一种用于测定代谢活性整个细胞内的酶的活性的测定化合物或其盐。 测定化合物包括离去基团和指示剂组。 离去基团选自包括氨基酸，肽，糖，硫酸盐，磷酸盐，酯，磷酸酯，核苷酸，多核苷酸，核酸，嘧啶，嘌呤，核苷，脂质及其混合物的组。 指示基团选自与离去基团连接时具有第一状态的化合物，当离去基团被酶从指示剂基团切割时的第二种状态。 优选地，指示剂化合物是这些化合物的罗丹明110，rhodol和荧光素以及类似物。 还公开了合成化合物的方法以及使用这些化合物测定酶活性的方法。