Department of Physiology, University of California Los Angeles, Los Angeles, California 90095-7327, United States.
Biochemistry. 2011 Dec 27;50(51):11009-14. doi: 10.1021/bi201592y. Epub 2011 Dec 1.
The sucrose permease (CscB) and lactose permease (LacY) of Escherichia coli belong to the oligosaccharide/H(+) symporter subfamily of the major facilitator superfamily, and both catalyze sugar/H(+) symport across the cytoplasmic membrane. Thus far, there is no common substrate for the two permeases; CscB transports sucrose, and LacY is highly specific for galactopyranosides. Determinants for CscB sugar specificity are unclear, but the structural organization of key residues involved in sugar binding appears to be similar in CscB and LacY. In this study, several sugars containing galactopyranosyl, glucopyranosyl, or fructofuranosyl moieties were tested for transport with cells overexpressing either CscB or LacY. CscB recognizes not only sucrose but also fructose and lactulose, but glucopyranosides are not transported and do not inhibit sucrose transport. The findings indicate that CscB exhibits practically no specificity with respect to the glucopyranosyl moiety of sucrose. Inhibition of sucrose transport by CscB tested with various fructofuranosides suggests that the C(3)-OH group of the fructofuranosyl ring may be important for recognition by CscB. Lactulose is readily transported by LacY, where specificity is directed toward the galactopyranosyl ring, and the affinity of LacY for lactulose is similar to that observed for lactose. The studies demonstrate that the substrate specificity of CscB is directed toward the fructofuranosyl moiety of the substrate, while the specificity of LacY is directed toward the galactopyranosyl moiety.
大肠杆菌的蔗糖通透酶(CscB)和乳糖通透酶(LacY)属于寡糖/H(+)协同转运蛋白超家族的成员,它们都能催化糖/H(+)协同跨细胞质膜转运。到目前为止,这两种通透酶没有共同的底物;CscB 转运蔗糖,而 LacY 对半乳糖吡喃糖苷高度特异。CscB 糖特异性的决定因素尚不清楚,但参与糖结合的关键残基的结构组织似乎在 CscB 和 LacY 中相似。在这项研究中,用过表达 CscB 或 LacY 的细胞测试了几种含有半乳糖吡喃基、葡萄糖吡喃基或果糖呋喃基部分的糖的转运。CscB 不仅识别蔗糖,还识别果糖和乳果糖,但葡萄糖苷不被转运,也不抑制蔗糖转运。这些发现表明,CscB 对蔗糖的葡萄糖吡喃基部分几乎没有特异性。用各种果糖呋喃糖苷测试 CscB 对蔗糖转运的抑制表明,果糖呋喃环的 C(3)-OH 基团可能对 CscB 的识别很重要。乳果糖很容易被 LacY 转运,LacY 的特异性针对半乳糖吡喃基环,LacY 对乳果糖的亲和力与对乳糖的亲和力相似。这些研究表明,CscB 的底物特异性针对底物的果糖呋喃基部分,而 LacY 的特异性针对半乳糖吡喃基部分。