Cytochemical characteristics of the golgi apparatus

Lectinocytochemistry provides a useful tool for localizing subcompartments of the complex reticular apparatus of Golgi. The technique is based on interactions of lectins with glycoconjugates present in the limiting membranes and luminal spaces of Golgi elements. Application of a series of lectins of different sugar specificities permits a differentiation between Golgi subcompartments containing glycoconjugates with different oligosaccharide side chains. These may be (a) different glycoconjugates or (b) glycoconjugates at different stages during synthesis or repair of their glycans. The lectinocytochemical studies with mannose-, glucose-, N-acetyl-glucosamine-, N-acetylgalactosamine-, galactose-, fucose-, and sialic acid-recognizing lectins revealed predominating patterns that labeled distinct, i.e., cis, medial, trans, and transmost, regions of the Golgi apparatus. A further refinement could be achieved by differential lectin-inhibition that enables a dissection of lectin binding reactions on the basis of their binding affinities. High-affinity binding reactions showed that subcompartments are not necessarily confined to one single Golgi subregion and may change their position from one to another subregion. Some of the patterns observed may be interpreted in relation to certain steps during synthesis and modifications of glycans.     

焙烤类谷物食品糖基化产物的定性、定量评价方法

糖基化产物与心血管疾病、糖尿病等多种慢性病的发生有关,因此对食品中糖基化产物含量的评价尤为重要。研究了不同酶解条件对结合态糖基化产物的释放,并且建立了以色度、吸光度、荧光强度等为考察指标综合评价不同反应途径的糖基化产物含量的体系。结果表明:分别采用E320μg/mL和640μg/mL酶解焙烤类谷类食物,40℃酶解36 h,结合态糖基化产物释放较为充分。谷物早餐中美拉德反应产物含量低于饼干和面包;荧光性晚期糖基化终产物含量大小依次为:饼干(1 890 AU/mg)面包(1 886 AU/mg)谷物早餐(1 678 AU/mg)。另外,早餐谷物中初期(A_(208))、中期糖基化产物(A_(360))显著高于饼干和面包(P0.05),而早餐谷物中期和末期糖基化反应产物(戊糖素、5-HMF和HMF)含量显著低于面包和饼干(P0.05)。     

Glycosylation,denaturation, and aggregation of soy proteins in defatted soy flakes flour: Influence of thermal and homogenization treatments

This article provides a systematic study of the impact of different thermal treatments (62 ± 2°C, without and with relative humidity control, 79%) on soy protein in defatted soy flour and their aqueous dispersions. The effect of dispersing treatments (magnetic stirring, high-speed, and high-pressure homogenization) on dispersions also was assessed. Changes in protein solubility (water and 0.2 g/100 g potassium hydroxide solution), apparent-reactive lysine content, urease and trypsin inhibitor activities, protein denaturation, and Fourier transform infrared spectra were studied. Glycosylation, aggregation, and denaturation of storage and biologically active soy proteins were observed in different degrees, being mainly promoted by the control of relative humidity and the dispersibility of the sample.     

糖基化终末产物对大鼠肾小管上皮细胞组织型转谷氨酰胺酶mRNA表达的影响

目的探讨晚期糖基化终末产物(Advanced glycosylation end products,AGEs)对大鼠肾小管上皮细胞NRK-52E组织型转谷氨酰胺酶(Tissue transglutanunase,tTG)mRNA表达及其对细胞外基质(Extracellular matrix,ECM)降解的影响。方法体外培养NRK-52E细胞,分别用体外合成的不同浓度的糖化牛血清白蛋白(AGE-BSA)及未经糖化的牛血清白蛋白(BSA)处理48h,ELISA法检测细胞培养上清中纤维连接蛋白(Fibronectin,FN)和Ⅳ型胶原蛋白(TypeⅣcollagen,ColⅣ)含量,RT-PCR检测细胞tTG mRNA的表达。结果与BSA组比较,50～200mg/LAGE-BSA组细胞上清中FN含量明显增加(P<0.01),25～100mg/L AGE-BSA组细胞上清中ColⅣ含量明显增加(P<0.05);AGE-BSA(50～400mg/L)可不同程度地刺激细胞tTG mRNA表达的增加(P<0.05);tTG mRNA表达增加与FN、ColⅣ分泌增多呈正相关(r值分别为0.911和0.872)。结论 AGEs能诱导大鼠近端肾小管上皮细胞tTG mRNA的表达,引起ECM主要成分FN和ColⅣ含量增加,提示tTG可能在AGEs引起的ECM积聚过程中,起到抑制ECM蛋白降解的作用,进而参与糖尿病肾病的病理过程。     

Recombinant expression and characterization of N-acetylglucosaminyltransferase I derived from Nicotiana tabacum

The C-terminal catalytic domain of tobacco N-acetylglucosaminyltransferase I fused to maltose-binding protein was produced in Escherichia coli as a soluble form with significant activity. The protein was affinity-purified using amylose resin, and its enzymatic properties were investigated, including its divalent cation requirements, optimal temperature, optimal pH, and substrate specificity.     

Tn and sialyl-Tn antigens,aberrant O-glycomics as human disease markers

In many different human disorders, the cellular glycome is altered. An interesting but poorly understood alteration occurs in the mucin-type O-glycome, in which there is aberrant expression of the truncated O-glycans Tn (GalNAcα1-Ser/Thr) and its sialylated version sialyl-Tn (STn) (Neu5Acα2,6GalNAcα1-Ser/Thr). Both Tn and STn are tumor-associated carbohydrate antigens and tumor biomarkers, since they are not expressed normally and appear early in tumorigenesis. Moreover, their expression is strongly associated with poor prognosis and tumor metastasis. The Tn and STn antigens are also expressed in other human diseases and disorders, such as Tn syndrome and IgA nephropathy. The major pathological mechanism for expression of the Tn and STn antigens is compromised T-synthase activity, resulting from alteration of the X-linked gene that encodes for Cosmc, a molecular chaperone specifically required for the correct folding of T-synthase to form active enzyme. This review will summarize our current understanding of the Tn and STn antigens in terms of their biochemistry and role in pathology.