Plasma alpha 1B-glycoprotein allele frequencies in Finns and Swedish Lapps: evidence for a new alpha 1B allele

A new allele (A1B*5) of human plasma alpha 1B-glycoprotein (alpha 1B) was reported. alpha 1B phenotyping was done by two-dimensional agarose gel (pH 5.4)-horizontal polyacrylamide gel (pH 9.0) electrophoresis followed by protein staining. The alpha 1B phenotypes 1-1, 1-2, 1-5 and 2-2 were observed in Finns and phenotypes 1-1, 1-2, 1-5 and 2-5 in Swedish Lapps. The respective frequencies of A1B*1, A1B*2 and A1B*5 were 0.9575, 0.0350, 0.0075 in Finns and 0.8922, 0.0653, 0.0425 in Swedish Lapps. The Swedish Lapps showed a higher degree of alpha 1B polymorphism (polymorphism information content = 0.19) than other Caucasian populations that have been studied.     

Donkey and horse alpha 1 B-glycoprotein: partial characterization and new alleles.

1. The donkey postalbumin protein has been shown to be the equivalent of human alpha 1 B-glycoprotein by protein immunoblotting and N-terminal amino acid sequence. 2. The horse A1B system (already identified as the homologue of human alpha 1 B-glycoprotein) and the donkey alpha 1 B-glycoprotein were characterized further for terminal sialic acid content, isoelectric point, amino acid composition and affinity for the dye-ligand, Cibacron Blue F3GA (known to bind human alpha 1 B-glycoprotein). 3. Two new alleles in the horse A1B system were found, bringing the total number of alleles to five. No polymorphism was found in the donkey alpha 1 B system. 4. As expected the first 20 N-terminal residues of the donkey and horse proteins are highly conserved with only two differences being found. 5. The polymorphism of the horse alpha 1 B-glycoproteins may be due in part to differing numbers of terminal sialic acid residues and the higher electrophoretic mobility of the donkey alpha 1 B-glycoprotein may be due in part to increased sialylation. 6. The horse and donkey alpha 1 B-glycoproteins exhibited differences in affinity for the dye-ligand, Cibacron Blue F3GA, with the donkey alpha 1 B-glycoprotein not being bound.     

Genetic polymorphism of alpha-2-HS-glycoprotein: four new alleles and allele frequencies in Japanese

alpha 2-HS-glycoprotein (AHSG) phenotyping was done in 655 Japanese from the Goto Islands, western Japan, using isoelectric focusing followed by immunoblotting. Four new AHSG alleles were encountered, AHSG*G1-G4, whose genetic transmissions were established in family studies. The allele frequencies were: AHSG*1 = 0.7221; AHSG*2 = 0.2748, and AHSG*G1-G4 = 0.0008, respectively.     

Two-dimensional electrophoresis of the plasma proteins of alpacas and llamas: genetic polymorphism of alpha 1B-glycoprotein and three other proteins

Plasma samples of alpacas and llamas were analysed by a simple method of two-dimensional (2-D) agarose gel (pH 8.6)-horizontal polyacrylamide gel (pH 9.0) electrophoresis, followed by general protein staining of gels. Genetic polymorphism in both species is described for alpha 1B-glycoprotein (alpha 1B) and three other unidentified proteins designated prealbumin (Pr), postalbumin 1 and 2 (Pa1 and Pa2). alpha 1B was identified by cross-reactivity with antisera for human and pig alpha 1B. Altogether, two alleles of Pr, two of Pa1, five of alpha 1B and three of Pa2 are described. Most of the alleles were present in alpacas and llamas. Alpacas showed a high degree of polymorphism at all four loci. Llamas showed considerable polymorphism at only the Pa1 and Pa2 loci. The theoretical probability of exclusion (PE) of an incorrectly assigned parent was estimated to be about 80% in each species by typing for the six polymorphic plasma proteins reported so far in these species. The given method of 2-D electrophoresis revealed no fixed differences in protein mobilities that discriminate between llamas and alpacas.     

Evidence for the presence of alpha 1B-glycoprotein in mammalian sera: immunoblotting studies

1. A monospecific antiserum to pig alpha 1B-glycoprotein (PO2) was produced in rabbits and was used to search for homologues of alpha 1B in sera of 41 mammalian species belonging to seven orders. 2. Specific reactions were detected in the sera of representatives of Insectivora, Primates, Carnivora, Proboscidea, Perissodactyla and Artiodactyla. No cross-reactions were observed in the sera of two species of Rodentia (mouse, rat). 3. Cross-reactions in the sera of Erinaceus europaeus, Homo sapiens and Macaca mulatta were rather weak; this indicates a greater structural difference between the alpha 1 B of Insectivora and Primates and that of the other mammalian orders. 4. Electrophoretic patterns of alpha 1 B were, in most cases, heterogeneous, the most heterogeneous being in ruminants. 5. Evidence was obtained that the alpha 1 B of sheep is identical with the earlier described (Juneja and Gahne (1980) Anim. Blood Grps Biochem. Genet. 11, 81-92.) polymorphic post-transferrin (Ptf).     

Pi Zpratt : A new alpha1-antitrypsin allele in an American negro family

Summary A low-power laser-UV microbeam of wave-length 257 nm was used for microirradiation of a small part of the nucleus of Chinese hamster cells. Following fixation in interphase or in the subsequent metaphase indirect immunofluorescent staining was performed with antiserum to photoproducts of DNA treated with far UV light.The results show that antibodies specific for UV-irradiated DNA can be used for a direct detection of laser-UV microirradiation-induced DNA photolesions. The potential usefulness of this method for investigation of the spatial arrangement of chromosomes in the interphase nucleus is discussed.     

Cysteine-rich secretory protein 3 is a ligand of alpha1B-glycoprotein in human plasma

Human cysteine-rich secretory protein 3 (CRISP-3; also known as SGP28) belongs to a family of closely related proteins found in mammals and reptiles. Some mammalian CRISPs are known to be involved in the process of reproduction, whereas some of the CRISPs from reptiles are neurotoxin-like substances found in lizard saliva or snake venom. Human CRISP-3 is present in exocrine secretions and in secretory granules of neutrophilic granulocytes and is believed to play a role in innate immunity. On the basis of the relatively high content of CRISP-3 in human plasma and the small size of the protein (28 kDa), we hypothesized that CRISP-3 in plasma was bound to another component. This was supported by size-exclusion chromatography and immunoprecipitation of plasma proteins. The binding partner was identified by mass spectrometry as alpha(1)B-glycoprotein (A1BG), which is a known plasma protein of unknown function and a member of the immunoglobulin superfamily. We demonstrate that CRISP-3 is a specific and high-affinity ligand of A1BG with a dissociation constant in the nanomolar range as evidenced by surface plasmon resonance. The A1BG-CRISP-3 complex is noncovalent with a 1:1 stoichiometry and is held together by strong electrostatic forces. Similar complexes have been described between toxins from snake venom and A1BG-like plasma proteins from opossum species. In these cases, complex formation inhibits the toxic effect of snake venom metalloproteinases or myotoxins and protects the animal from envenomation. We suggest that the A1BG-CRISP-3 complex displays a similar function in protecting the circulation from a potentially harmful effect of free CRISP-3.     

The ESD polymorphism: Further studies of the ESD2 and ESD5 allele products

Summary Electrofocusing and agarose electrophoresis techniques both reveal polymorphism of ESD 2, which may be subdivided into two different proteins, coded for by genes allelic to ESD ^*1. After agarose electrophoresis, ESD 2 is slightly more anodally located than ESD 5, while the latter is considerably more acidic as revealed by electrofocusing in polyacrylamide gel slabs. Family studies have confirmed that each of the allele products behave as Mendelian characters: and the gene frequencies in a Norwegian population material are about 0.08 and 0.02 for the ESD ^*2 and ESD ^*5 alleles, respectively.     

Genotype and allele frequencies of divalent metal transporter 1 polymorphism in Turkish population

Divalent metal transporter 1 (DMT1 also known as DCT1, NRAMP2 or SCL11A2) is a membrane-bound divalent metal transporter which is conserved from prokaryotes to higher eukaryotes. It has been postulated to play important roles in intestinal iron absorption at the brush border of duodenal enterocytes, erythroid iron utilization, hepatic iron accumulation, placental iron transfer, and other processes. DMT1 gene which contains at least four isoforms (1A/+IRE, 1A/-IRE, 2/+IRE and 2/-IRE) is located on chromosome 12q13 in human. DMT1 mediates the transport of a wide range of metals, including the essential metals Fe2+, Zn2+, Mn2+, Cu2+, Co2+, Ni2+ and toxic metals such as Cd2+ and Pb2+. The intention of this study is to determine that IVS4+44C/A single nucleotide polymorphism in DMT1 gene of Turkish population. For this purpose blood samples from 192 female and 192 male volunteers were analyzed. DMT1 gene was amplified with the polymerase chain reaction-restriction fragment length polymorphism technique and 351 bp oligonucleotide was produced. The amplified oligonucleotides were cut with MnlI restriction enzyme according to their polymorphic characteristics. Digested and undigested products were separated on a 2% agarose gel electrophoresis, visualized by ethidium bromide staining under an ultraviolet illuminator. The genotype frequencies of DMT1 IVS4+44C/A polymorphism were determined as 47.9% for CC, 40.1% for AC and 12.0% for AA genotypes. The frequency of the C allele was found to as 68.0% and of the A allele as 32.0%. The genotype frequencies were consistent with Hardy-Weinberg equilibrium (χ2=2.394; Exact P=0.128).     

PiT: A new allele in the alpha1-antitrypsin system

Summary In a genetic investigation of the population in Hessen, Germany, we found a family with a new, rare allele in the Pi system (₁-antitrypsin). According to electrophoretic analysis and isoelectric focusing patterns, it is designated Pi ^T. A pedigree study suggests autosomal codominant inheritance. The serum concentration of six heterozygous carriers of this allele (phenotype M1T or M2T) revealed normal ₁-antitrypsin levels.     

Frequencies of plasma protease inhibitor alleles in Australian horse breeds and the recognition of two new alleles

Investigation of the plasma protease inhibitor system (Pi) in the Arabian and quarter horse breeds and re-examination of the standardbred breed resulted in the recognition of two new Pi alleles, designated E and L2. PiE is rare and has been found in only three quarter horses. In contrast, PiL2 is relatively common in the standardbred (0.107) and allowed subdivision of PiL into PiL and PiL2. Splitting of PiL resulted in an exclusion probability (PE) of 0.649 for the standardbred Pi system. Frequencies of the Pi genes have now been determined for four breeds (thoroughbred, standardbred, quarter horse and Arabian) of horses in Australia.     

A new allele of human alpha1-antitrypsin: PiNhampton.

A new allele of alpha1AT is described. By isoelectric focusing, the microheterogeneous pattern of the variant was similar to but more cathodal than that of Pi N. This allele has therefore been tentatively designated PiNhampton(Nham). Further examination revealed that the minor bands of Nham are indistinguishable from the major bands of Z by isoelectric focusing, and a careful family study was necessary to clearly define the proband's phenotype. Pi Nham was found in association with M1, S, and Z, but to date its possession is not apparently related to clinical disorders or reduced serum levels of alpha1AT.     

Two-dimensional electrophoresis of the plasma proteins of alpacas and llamas: genetic polymorphism of α1B-glycoprotein and three other proteins

Summary. Plasma samples of alpacas and llamas were analysed by a simple method of two-dimensional (2-D) agarose gel (pH 8.6)-horizontal polyacrylamide gel (pH 9.0) electrophoresis, followed by general protein staining of gels. Genetic polymorphism in both species is described for α 1 B-glycoprotein (α 1 B) and three other unidentified proteins designated prealbumin (Pr), postalbumin 1 and 2 (Pal and Pa2). α 1 B was identified by cross-reactivity with antisera for human and pig α 1 B. Altogether, two alleles of Pr, two of Pa1, five of α 1 B and three of Pa2 are described. Most of the alleles were present in alpacas and llamas. Alpacas showed a high degree of polymorphism at all four loci. Llamas showed considerable polymorphism at only the Pa1 and Pa2 loci. The theoretical probability of exclusion (P e ) of an incorrectly assigned parent was estimated to be about 80% in each species by typing for the six polymorphic plasma proteins reported so far in these species. The given method of 2-D electrophoresis revealed no fixed differences in protein mobilities that discriminate between llamas and alpacas.     

C1R subcomponent polymorphism in Japanese: description of a new allele

The polymorphism of C1R was investigated in 570 unrelated Japanese individuals using isoelectric focusing and immunoblotting. A total of 11 different C1R phenotypes including a new pattern designated C1R 11-1 were observed. The allele frequencies were C1R*1 = 0.4561, C1R*2 = 0.3377, C1R*5 = 0.1956, C1R*8 = 0.0088 and C1R*R (C1R*9 and C1R*11) = 0.0018. The population data fitted the Hardy-Weinberg equilibrium. The C1R polymorphism in Japanese was shown to be controlled by 3 common alleles, C1R*1, C1R*2 and C1R*5, as compared to Caucasians where only the former 2 are present commonly. This complement system can be a useful genetic marker for anthropological studies.     

Pig plasma postalbumin-2 (alpha 1B-glycoprotein): isolation, partial characterization and immunological cross-reactivity with other mammalian sera

1. Components of pig plasma postalbumin-2 (PO2) protein, after rivanol-ammonium sulphate fractionation of plasma, were separated from other proteins by an easy and rapid method of horizontal double-one dimensional IPG-PAGE. The protein was recovered from polyacrylamide gel by combination of electrophoresis and isoelectric focusing. 2. The mol. wt of PO2 was estimated to be 68,000, using SDS-PAGE. 3. Amino acid and carbohydrate compositions of PO2 were very similar to those of human plasma alpha 1B-glycoprotein (alpha 1B), confirming that PO2 is the porcine homologue of human alpha 1B. 4. Neuraminidase treatment resulted in a decrease of electrophoretic migration velocity of all four studied components of PO2. 5. Homologous proteins to pig PO2 (alpha 1B) were observed, not only in human plasma but also in plasma of dog, horse and rabbit, by immunoblotting.     

Apolipoprotein A-IV polymorphism in the Finnish population: gene frequencies and description of a rare allele

The apolipoprotein A-IV (apoA-IV) allele frequencies were determined in 387 adult Finns by immunoblotting after isoelectric focusing of serum. The gene frequencies were: A-IV1 = 0.942 and A-IV2 = 0.058. The phenotypes of 147 mother-child pairs studied were in accordance with the two allelic modes of inheritance. In 2 subjects, a rare apoA-IV variant was found.     

Salivary protein polymorphism in Kenya: evidence for a new AMY1 allele

Salivary protein polymorphism was studied in 200 schoolboys, mainly Kisii and Luo from Kenya, East Africa. The frequencies of PR, PA, DB, PB and AMY1 genes were as follows: PR*1: 0.66, PA*(+): 0.18, DB*(+): 0.55, PB*2: 0.12, AMY1*A2: 0.008, AMY1*E: 0.03. These frequencies were compared with other population data, in particular from West African and US Negroes. The most interesting finding with respect to the gene frequencies is the low PB*2 frequency and the absence of AMY1*3 in Kenya. Furthermore, a new phenotype in the AMY1 system was described which suggests the presence of an allele with an estimated frequency of 0.02.     

Further studies on proteinases and alpha 2-macroglobulin activity in diabetic plasma.

Loss of chymotrypsin binding capacity of alpha 2-macroglobulin in diabetic plasma on in vitro incubation, could be partially prevented by phenylmethyl sulphonyl fluoride and pepstatin A. Prior ten-fold dilution of plasma with 0.02 M phosphate buffer (pH 7.0) completely arrested the process. The phenomenon could not be reactivated by Ca2+, lecithin or bovine serum albumin. Diabetic plasma, like normal plasma, exhibited maximal hydrolytic activities on H-D-Pro-Phe-Arg-p-nitroanilide, H-D-Val-Leu-Arg-p-nitroanilide and H-D-Ile-Pro-Arg-p-nitroanilide. The hydrolytic activities were not significantly diminished on incubation of plasma at 37 degrees C for 12 hr, unlike alpha 2-macroglobulin activity. On gel chromatography on Sephadex G-200, part of the proteolytic activity in diabetic plasma coeluted with alpha 2-macroglobulin in the VO region. A second activity peak (absent in normal plasma) was eluted with a Ve/V0 value of 1.40. Possible role of free proteinases in diabetic plasma in the inactivation of alpha 2-macroglobulin is discussed.     

Identification and DNA sequence analysis of 15 new alpha 1-antitrypsin variants, including two PI*Q0 alleles and one deficient PI*M allele.

We have investigated the molecular basis of 15 new alpha 1-antitrypsin (alpha 1AT) variants. Phenotyping by isoelectric focusing (IEF) was used as a screening method to detect alpha 1AT variants at the protein level. Genotyping was then performed by sequence analysis of all coding exons, exon-intron junctions, and the hepatocyte-specific promoter region including exon Ic. Three of these rare variants are alleles of clinical relevance, associated with undetectable or very low serum levels of alpha 1AT:the PI*Q0saarbruecken allele generated by a 1-bp C-nucleotide insertion within a stretch of seven cytosines spanning residues 360-362, resulting in a 3' frameshift and the acquisition of a stop codon at residue 376; a point mutation in the PI*Q0lisbon allele, resulting in a single amino acid substitution Thr68(ACC)-->Ile(ATC); and an in-frame trinucleotide deletion delta Phe51 (TTC) in the highly deficient PI*Mpalermo allele. The remaining 12 alleles are associated with normal alpha 1AT serum levels and are characterized by point mutations causing single amino acid substitutions in all but one case. This exception is a silent mutation, which does not affect the amino acid sequence. The limitation of IEF compared with DNA sequence analysis, for identification of new variants, their generation by mutagenesis, and the clinical relevance of the three deficiency alleles are discussed.