Bach G, Kohn G, Lasch E E, El Massri M, Ornoy A, Sekeles E, Legum C, Cohen M M
Pediatr Res. 1978 Oct;12(10):1010-5. doi: 10.1203/00006450-197810000-00012.
A partial deficiency of alpha-mannosidase was found in cultured skin fibroblasts, serum, and extracts of leukoytes in two siblings with mild mental retardation, delayed speech, a suggestion of coarse or full facies, and limited mobility of the large joints. All other lysosomal enzymes tested were within the normal range. Their father demonstrated intermediate alpha-mannosidase activity. The addition of 2 mM Zn++ caused a 40% increase of the alpha-mannosidase activity in cell extracts of both patients and control subjects. pH profiles and Cellogel electrophoresis of the patients' cells indicated 20% residual activity of the acidic alpha-mannosidase isoenzyme (pH optimum at 4.0), whereas the activity of the isozyme with pH optimum of 6.0 was normal. Increasing substrate concentration (1--10 mM) demonstrated a 4 to 5-fold increase in the apparent Km of the acidic alpha-mannosidase in the patients' fibroblasts. This residual activity, however, was apparently not sufficient for the normal catabolism of mannose-containing molecules, since electron microscopic examination of the cultured fibroblasts demonstrated numerous lysosomal storage bodies.
在两名患有轻度智力迟钝、语言发育迟缓、面部有粗糙或丰满迹象且大关节活动受限的同胞兄妹的培养皮肤成纤维细胞、血清和白细胞提取物中,发现了α-甘露糖苷酶部分缺乏。所检测的所有其他溶酶体酶均在正常范围内。他们的父亲表现出中等水平的α-甘露糖苷酶活性。添加2 mM Zn++可使患者和对照受试者细胞提取物中的α-甘露糖苷酶活性增加40%。患者细胞的pH曲线和Cellogel电泳表明,酸性α-甘露糖苷酶同工酶(最适pH为4.0)有20%的残余活性,而最适pH为6.0的同工酶活性正常。增加底物浓度(1 - 10 mM)表明患者成纤维细胞中酸性α-甘露糖苷酶的表观Km增加了4至5倍。然而,这种残余活性显然不足以进行含甘露糖分子的正常分解代谢,因为对培养的成纤维细胞进行电子显微镜检查发现了大量溶酶体储存体。