中国广东人红细胞PGM_1亚型遗传性多态现象

采用薄层聚丙烯酰胺凝胶等电聚焦技术,调查了中国(广东)406名无亲缘关系的正常人红细胞磷酸葡萄糖变位酶-1(PGM_1)亚型的遗传多态性。除了常见的10种亚型外,还发现了由一个新的变异型等位基因和常见的4个等位基因杂合产生的9例变异型。PGM_1位点的等位基因频率PGM_1~(1+)、PGM_1~(1-)、PGM_1~(2+)、PGM-1~(2-)和PGM_1~(V丰)(变异型等位基因)分别为0.5973、0.1256、0.1724、0.0936和0.0111;群体处于Hardy-Weinberg式平衡状态。变异型等位基因以多态频率出现,可能成为该群体的一个重要的遗传性特征。     

我国几个民族中毛发根葡糖磷酸变位酶—1的亚型分布

本文报道了利用等电聚焦的方法,从毛发根鞘细胞分析了我国汉、苗、土家、撒拉、土、达斡尔和赫哲等民族的葡糖磷酸变位酶-1(PGM_1)10亚型的分布,并且与其他人群的基因频率作了比较。     

上海地区胃癌病人运铁蛋白(Tf)、组特异性成分(Gc)、α_1-抗胰蛋白酶(α_1-AT)亚型分布的研究

用薄层聚丙烯酰胺凝胶等电聚焦电泳分析了上海地区202例汉族胃癌病人及202例正常对照组的运铁聋白(Tf)和α_1-抗胰蛋白酶(α_1-AT)亚型的分布,发现胃癌组Tfc_1c_1纯合子频率(0.3713)和Tfc_1基因频率(0.5718)显著低于对照组(分别为0.5149和0.6782),均为p<0.01,胃癌组Tfc_2c_2纯合子频率(0.2228)和Tfc_2基因频率(0.4019)显著高于对照组(分别为0.1436,p<0.05和0.2970,p<0.01),胃癌组和对照组的α_1-抗胰蛋白酶亚型分布无显著差异。用薄层聚丙烯酰胺等电聚焦结合免疫固定分析了上海地区200例汉族胃癌病人和200例正常对照组的组特异性成分(Gc)亚型分布,发现胃癌组Go1F表型频率(0.22)和Gc1F基因频率(0.4375)均显著高于正常对照组(分别为0.14和0.3600,均为p<0.05)。     

云南省元江县白族人红细胞EsD和PGM_1分型的调查

采用淀粉/琼脂糖混合凝胶同步电泳法,对云南省元江县126例白族人红细胞EsD和PGM_1作了分型调查,计算出这两种红细胞同乙酶的基因频率及识别能力,结果表明云南省元江县白族与上述两地居民(汉族)的红细胞EsD和PGM_1表型分布无显著性差异。     

中国20个民族中PGM_1及其亚型EsD、GLO1、AK、ADA和6-PGD酶型分布调查

调查了汉族、鄂伦春、赫哲、朝鲜、蒙古、羌、土家、苗、侗、畲、壮、纳西、傈僳、白、彝、景颇、哈尼、傣、维吾尔和塔吉克等20个民族的PGM_1及其亚型,EsD、GLO_1、AK、ADA和6-PGD等酶型的分布及基因频率。PGM_1及其亚型、EsD和GLO_1在中国各民族中是分布较好的,个人识别能力较高的酶。有12个民族查出有PQM_1~6基因,壮族的频率最高,PGM_1 6-1表型达4.15％。对在4174份血样中所检出的带有PGM_1~6基因的68份血样做亚型分析,在凝胶上PGM_1~6谱带均在同一位置上。EsD_1基因频率的总趋向是北方各民族高于南方。哈尼、傈僳、傣、纳西、畲、壮、侗和苗等民族EsD2-2表型达15％以上,哈尼族高达32.4％。GLO1~1基因频率塔吉克和维吾尔族为0.2927和0.2112,羌族为0.0583,其它各族在0.0714—0.1527。各民族AK~1、ADA和6-PGD~(?)基因频率均甚高。     

中国人群葡萄糖磷酸变位酶基因频率测定及PGM18-1表型检出

葡萄搪磷酸变位酶(phosphoglucomutase, PGM）是糖元代谢中十分重要的酶,它可逆地 催化下列反应：a-D-葡萄搪-1一磷酸二兰生全 G-1,6-2P a-D-葡萄糖-6-磷酸盐。该酶是由定位于1,4, 6染色单体上PGM PGM PGM,位点上的 等位基因所控制。PGM 广泛分布于体内各种 组织中,其中80-90多为PGM,的基因产物。 红细胞中PGM,编码的同工酶活性与PGM, 活性各占50多,在红细胞中检测不出PG嶙基 因的活性产物。 1964年Spencer等人(47先后证明PGM：具 有多态性。采用淀粉凝胶法检测出PGM, 1, PGM, 2-1和PGM, 2三种遗传表型,它们是 由第1对染色体上PGM,位点上的两个常见 的等位基因PGMi和PGM,所决定。高分辨 的等电聚焦电泳可检出PG城有10种亚型, 是由PGM,位点上的4个等位基因PGMi十、 PGM犷、PG叫十和PG域一所编码121。本文报 告我们应用国产淀粉,采用混合淀粉凝胶方法 测定中国（北京地区）人群的基因频率分布,同 时报道对PGM, 8-1表型的检出。     

琼脂糖平板制备聚焦电泳法进一步提纯胸腺素组分五

用琼脂糖平板等电聚焦电泳法,由胸腺素组分五中分离出三种在聚焦电泳谱上是单一谱线的多肽成份——CP1、CP2和CP3,等电点分别为4.3、4.9和5.6。测定了这些多肽对脐带血中淋巴细胞形成羊红细胞玫瑰花的影响。与对照相比,CP1(2微克/0.6毫升),和CP3(0.2-2微克/毫升)分别在统计学上呈显著和非常显著差异。在相同测定条件下,这三种多肽成份的活性均高于化学合成的胸腺多肽——胸腺素α_1。     

三种分析水稻F1代幼苗生长过程中酯酶同工酶变化的电泳方法比较

利用聚酯胶片作为凝胶支持物的超薄等电聚焦电泳(UTLIEF)研究水稻F1种子萌发过程中的酯酶同工酶多态性变化的结果表明,UTLIEF相比非变性不连续聚丙烯酰胺电泳(native-PAGE)和常规等电聚焦电泳(IEF)得到的酯酶同工酶图谱更清晰,酶带数目和强弱的多态性变化更高,用pH 2～9的两性电解质时等电聚焦的电泳效果较好.     

大熊猫子宫钙调素的分离纯化和性质的研究

以大熊猫子宫为材料分离纯化了钙调素(Calmodulin,CaM),经SDS-PAGE,PAGE和等电聚焦电泳鉴定,表现均一。分子量为18800道尔顿,等电点为3.6。该蛋白质分子的N-末端为封闭的。大熊猫子宫钙调素具有其它来源钙调素所特有的一些性质。对环核苷酸磷酸二酯酶有明显的激活作用,还发现对超氧化物歧化酶也有一定的激活作用。电泳行为受Ca~(2+)影响而出现特征性电泳改变,在含有Ca~(2+)的SDS凝胶电泳中,电泳速度比EGTA存在对略快,在PAGE中,有Ca~(2+)比无Ca~(2+)对电泳速度略慢。大熊猫子宫钙调素的氨基酸组成中,Phe/Tyr为8:2,可观察到钙调素特征性紫外吸收光谱。     

野生纤毛虫同工酶微量等电聚焦分析探索

微量电泳是用来分离和表征提取于少量生物材料中的核酸和蛋白质的分析技术。作者以缘毛目螅状独缩虫(Carchesium polypinum Linne,1758)为材料,用微量等电聚焦(Microisoelectric focussing)探索了分析野生纤毛虫同工酶的可行性。实验结果表明:(1) 样品量小至两个群体螅状独缩虫(约200个个体)即可进行酯酶同工酶微量等电聚焦分析;(2) 自野外采集材料中立即分离制备的螅状独缩虫匀浆上清液的酯酶同工酶酶谱与实验室内放置10h后分离制备的酶谱几乎一致。因此,野生纤毛虫同工酶可用微量等电聚焦进行分析。     

Isoelectric focusing studies of human red cell PGM1 in Japanese, with special reference to the characterization of PGM17

The distribution of phosphoglucomutase (PGM1) subtypes in human red cells was determined by isoelectric focusing in 218 Japanese samples. Nine common phenotypes were observed corresponding to the following frequencies of the four alleles at the PGM1 locus: PGM11+ 0.6560, PGM11- 0.1170, PGM12+ 0.1674 and PGM12- 0.0505. In addition, a characterization of the PGM17 allele was performed. Our results obtained in the present study revealed the possibility that the PGM17 allele may be differentiated in the two alleles of PGM17+ and PGM17- through an investigation of isoelectric focusing.     

PGM1 subtype polymorphism in 14 endogamous Dravidian-speaking populations of South India

Red cell hemolysates from 1,004 persons belonging to 14 population groups drawn from four South Indian states, Andhra Pradesh, Tamil Nadu, Karnataka, and Kerala, were tested for PGM1 subtypes. The groups are characterized by a high frequency of phenotype 1+1+ (range 36.98-71.64%) and the allele 1+ (range 60-79%). The groups exhibit marked heterogeneity for PGM1 locus. The results show a clear demarcation between tribes and Brahmin groups.     

Genetic studies of phosphoglucomutase-1 (PGM1) subtypes: population aspects]

Distribution of the subtypes and gene frequencies of phosphoglucomutase-1 among some populations of Buryats, Kirghizes of the Pamir and Russians of Moscow district was analysed. The frequencies of PGM1 genes vary in Buryats being PGM1+(1) 0.647-0.743, PGM1-(1)-0.100-0.132, PGM2+(1)-0.122-0.199 and PGM2-(1)-0.007-0.037. Following frequencies of PGM1 genes were established for Kirghizes: PGM1+(1) = 0.614, PGM1-(1) = 0.114, PGM2+(1) = 0.217 and PGM2-(1) = 0.054; in Russian populations the frequencies were: PGM1+(1) = 0.578, PGM1-(1) = 0.110, PGM2+(1) = 0.253 and PGM2-(1) = 0.059. Peculiarities of PGM1 polymorphism in the USSR and all over the world were analysed. Parallel biodemographic investigations in Buryat population demonstrated differences in intensities of selection, related to concrete PGM genotypes.     

To the research of the gene pool of the Gagauz population of Moldavia

Population genetic data on Gagauzes from Moldavia are reported here for the first time. AB0 and Rhesus blood groups, serum protein group (HP, TF, GC) and the red cell enzyme polymorphism PGM1 were determined in 190 Gagauzes. In addition to this the ability to taste PTC was tested. The following allele frequencies were found: AB0*0 = 0.5241, AB0*A = 0.3279, AB0*B = 0.1480; RH*D = 0.6083, RH*d = 0.3917; HP*1 = 0.3544, HP*2 = 0.6456; TF*C1 = 0.7472, TF*C2 = 0.1770, TF*C3 = 0.0730, TF*B = 0.0028; GC*1F = 0.1025, GC*1S = 0.5932, GC*2 = 0.3043; PGM*1+ = 0.5932; PGM*1- = 0.1000, PGM*2+ = 0.2607, PGM*2- = 0.1107. The frequency of the PTC*T allele was found to be 0.5298. These frequencies and genetic distance analyses show that the gene pool of the Gagauzes is similar to that of neighbouring southeastern European populations.     

Isoelectric focusing of human red cell phosphoglucomutase (PGM1). Phenotype distribution in the population of Tuscany and two hereditary variants

Summary Phosphoglucomutase (PGM₁) phenotypes were determined in a population sample of Tuscany, Italy, by isoelectric focusing. The frequencies observed for the four alleles are: PGM ₁ ¹⁺ =0.6012, PGM ₁ ^1- =0.1059, PGM ₁ ²⁺ =0.2495, PGM ₁ ^2- =0.0434. Two variants were detected and it was possible to study the parentage of both of them. The pedigree of the propositus of the first variant shows that the variant occurs in combination with the common alleles PGM₁ 1+ and PGM₁ 2+ and that it has an autosomal dominant inheritance. The second variant has been shown to be a product of the PGM₂ locus, although its PAGIF pattern is included between 2- and 1+ isoenzymes.     

Purification, characterization, and molecular cloning of a 60-kDa phosphoprotein in rabbit skeletal sarcoplasmic reticulum which is an isoform of phosphoglucomutase.

A 60-kDa substrate of calmodulin-dependent protein kinase in rabbit "heavy" skeletal sarcoplasmic reticulum (SR) was characterized by purification and cDNA cloning. Purification was achieved by column chromatography using DEAE-Sephacel, heparin-agarose, and hydroxylapatite in 0.5% 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonic acid (CHAPS). Analyses of amino acid sequence and composition indicated that the CHAPS-soluble 60-kDa protein is an isoform of phosphoglucomutase (PGM). cDNAs encoding two isoforms of PGM were isolated from rabbit skeletal muscles. The translated amino acid sequences show that the isoforms, PGM1 and PGM2, differ in the N-terminal 77 amino acids and that PGM2 is identical to the 60-kDa protein in the SR. Northern blot analysis showed that the size of the mRNA encoding PGM2 is 2.4 kilobases. The PGM enzyme activity was markedly inhibited in SR membranes, while perturbation of the membranes with CHAPS or guanidine-HCl recovered the enzyme activity. KCl (0.15-1 M) led to a partial recovery of the enzyme activity suggesting that the charge interaction is not the primary force for PGM-SR interaction. PGM is localized in the heavy fraction of SR, where calsequestrin and Ca2+ release channel are enriched. Our results demonstrate that an isoform of PGM localized in junctional skeletal SR is the 60-kDa substrate of calmodulin-dependent protein kinase.     

Catalytic properties of phosphoglucomutase from pea chloroplasts.

Electrophoretically homogeneous phosphoglucomutase (PGM) with specific activity of 3.6 units/mg protein was isolated from pea (Pisum sativum L.) chloroplasts. The molecular mass of this PGM determined by gel-filtration is 125 +/- 4 kD. According to SDS-PAGE, the molecular mass of subunits is 65 +/- 3 kD. The Km for glucose-1-phosphate is 18.0 +/- 0.5 microM, and for glucose-1, 6-diphosphate it is 33 +/- 0.7 microM. At glucose-1-phosphate and glucose-1,6-diphosphate concentrations above 0.5 and 0.2 mM, respectively, substrate inhibition is observed. The enzyme has optimum activity at pH 7.9 and 35 degrees C. Mg2+ activates the PGM. Mn2+ activates the enzyme at concentrations below 0.2 mM, while higher concentrations have an inhibitory effect. The activity of the PGM is affected by 6-phosphogluconate, fructose-6-phosphate, NAD+, ATP, ADP, citrate, and isocitrate.     

A highly active phosphoglucomutase from Clostridium thermocellum: cloning, purification, characterization and enhanced thermostability

Aims: Discovery and utilization of highly active and thermostable phosphoglucomutase (PGM) would be vital for biocatalysis mediated by multiple enzymes, for example, high‐yield production of enzymatic hydrogen. Methods and Results: The thermophilic cellulolytic bacterium Clostridium thermocellum was hypothesized to have a very active PGM because of its key role in microbial cellulose utilization. The Cl. thermocellum ORF Cthe1265 encoding a putative PGM was cloned and expressed in Escherichia coli. The purified enzyme appeared to be a monomer with an estimated molecular weight of 64·9 kDa. This enzyme was found to be a dual‐specificity enzyme – PGM/phosphomannomutase (PMM). Mg²⁺ and Mn²⁺ were activators. Ser144 was identified as an essential catalytic residue through site‐directed mutagenesis. The k_cat and K_m of PGM were 190 s^?1 and 0·41 mmol l^?1 on glucose‐1‐phosphate and 59 s^?1 and 0·44 mmol l^?1 on mannose‐1‐phosphate, respectively, at 60°C. Thermostability of PGM at a low concentration (2 nmol l^?1, 100 U l^?1) was enhanced by 12‐fold (i.e. t_1/2 = 72 h) at 60°C with addition of bovine serum albumin, Triton X‐100, Mg²⁺and Mn²⁺. Conclusions: The ORF Cthe1265 was confirmed to encode a PGM with PMM activity. This enzyme was the most active PGM reported. Significance and Impact of the Study: This highly active PGM with enhanced thermostability would be an important building block for in vitro synthetic biology projects (complicated biotransformation mediated by multiple enzymes in one pot).