啮齿动物肝脏α-L-岩藻糖苷酶的同工型、动力学及免疫化学特性

Isoform, kinetic and immunochemical characterization of rodent liver alpha-L-fucosidases.

作者信息

Johnson S W, Alhadeff J A

机构信息

Department of Chemistry, Lehigh University, Bethlehem, PA 18015.

出版信息

Comp Biochem Physiol B. 1990;97(4):713-7. doi: 10.1016/0305-0491(90)90112-7.

DOI:10.1016/0305-0491(90)90112-7

PMID:2085955

Abstract

alpha-L-Fucosidase from hamster and six inbred mouse strains contains two to three unique basic isoelectric forms (above pI 7.0) in addition to the usual acidic and neutral isoforms from pI 4-7. Rat liver alpha-L-fucosidase contains multiple isoforms between pI values of 4.0 and 7.3 whereas guinea pig liver alpha-L-fucosidase exhibits a single broad isoform at pI 5.3. 2. All the alpha-L-fucosidases have similar KM values (0.05-0.12 mM) for 4-methylumbelliferyl-alpha-L-fucopyranoside but pH-activity curves which are significantly different in optima and per cent of optimal activity in the acid region. 3. Double-antibody immunoprecipitation experiments indicate that rodent liver alpha-L-fucosidases crossreact to varying extents with polyclonal antibody against human liver alpha-L-fucosidase. 4. Hamster, guinea pig and mouse liver alpha-L-fucosidases exhibit significantly less binding than human and rat liver fucosidases to the agarose-epsilon-aminocaproylfucosamine affinity resin.

摘要

仓鼠和六种近交系小鼠的α-L-岩藻糖苷酶除了具有通常的pH值为4-7的酸性和中性同工型外，还含有两到三种独特的碱性等电形式（pH值高于7.0）。大鼠肝脏α-L-岩藻糖苷酶在pH值4.0至7.3之间含有多种同工型，而豚鼠肝脏α-L-岩藻糖苷酶在pH值5.3处表现为单一的宽同工型。2. 所有的α-L-岩藻糖苷酶对4-甲基伞形酮基-α-L-岩藻吡喃糖苷的米氏常数（KM）相似（0.05-0.12 mM），但pH-活性曲线在最佳pH值和酸性区域的最佳活性百分比方面有显著差异。3. 双抗体免疫沉淀实验表明，啮齿动物肝脏α-L-岩藻糖苷酶与抗人肝脏α-L-岩藻糖苷酶的多克隆抗体有不同程度的交叉反应。4. 仓鼠、豚鼠和小鼠肝脏α-L-岩藻糖苷酶与人及大鼠肝脏岩藻糖苷酶相比，与琼脂糖-ε-氨基己酰岩藻糖胺亲和树脂的结合明显较少。