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1.
BMC Genet ; 16: 136, 2015 Dec 02.
Article in English | MEDLINE | ID: mdl-26628212

ABSTRACT

BACKGROUND: Located in the Pacific Ocean between Australia and New Zealand, the unique population isolate of Norfolk Island has been shown to exhibit increased prevalence of metabolic disorders (type-2 diabetes, cardiovascular disease) compared to mainland Australia. We investigated this well-established genetic isolate, utilising its unique genomic structure to increase the ability to detect related genetic markers. A pedigree-based genome-wide association study of 16 routinely collected blood-based clinical traits in 382 Norfolk Island individuals was performed. RESULTS: A striking association peak was located at chromosome 2q37.1 for both total bilirubin and direct bilirubin, with 29 SNPs reaching statistical significance (P < 1.84 × 10(-7)). Strong linkage disequilibrium was observed across a 200 kb region spanning the UDP-glucuronosyltransferase family, including UGT1A1, an enzyme known to metabolise bilirubin. Given the epidemiological literature suggesting negative association between CVD-risk and serum bilirubin we further explored potential associations using stepwise multivariate regression, revealing significant association between direct bilirubin concentration and type-2 diabetes risk. In the Norfolk Island cohort increased direct bilirubin was associated with a 28% reduction in type-2 diabetes risk (OR: 0.72, 95% CI: 0.57-0.91, P = 0.005). When adjusted for genotypic effects the overall model was validated, with the adjusted model predicting a 30% reduction in type-2 diabetes risk with increasing direct bilirubin concentrations (OR: 0.70, 95% CI: 0.53-0.89, P = 0.0001). CONCLUSIONS: In summary, a pedigree-based GWAS of blood-based clinical traits in the Norfolk Island population has identified variants within the UDPGT family directly associated with serum bilirubin levels, which is in turn implicated with reduced risk of developing type-2 diabetes within this population.


Subject(s)
Bilirubin/blood , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/genetics , Genetic Predisposition to Disease , Glucuronosyltransferase/genetics , Haplotypes/genetics , Alleles , Base Sequence , Cardiovascular Diseases/complications , Cardiovascular Diseases/genetics , Chromosomes, Human, Pair 2/genetics , Diabetes Mellitus, Type 2/enzymology , Genes, Recessive , Genome-Wide Association Study , Humans , Inheritance Patterns/genetics , Linkage Disequilibrium , Melanesia , Metabolic Syndrome/complications , Metabolic Syndrome/genetics , Molecular Sequence Annotation , Molecular Sequence Data , Polymorphism, Single Nucleotide/genetics , Risk Factors
2.
Transfus Med ; 25(5): 326-32, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26132409

ABSTRACT

OBJECTIVES: The major aims of this study are to characterise and compile allelic data of human platelet antigen (HPA)-1 to -6 and -15 systems in five Malay sub-ethnic groups in Peninsular Malaysia. BACKGROUND: HPAs are polymorphic glycoproteins expressed on the surface of platelet membranes and are genetically differentiated across ethnogeographically unrelated populations. METHODS: Blood samples were obtained with informed consent from 192 volunteers: Banjar (n = 30), Bugis (n = 37), Champa (n = 51), Jawa (n = 39) and Kelantan (n = 35). Genotyping was done using polymerase chain reaction-sequence specific primer method. RESULTS: In general, frequencies of HPAs in the Malay sub-ethnic groups are more similar to those in Asian populations compared with other more distinct populations such as Indians, Australian Aborigines and Europeans. CONCLUSIONS: This study provides the first HPA datasets for the selected Malay sub-ethnic groups. Subsequent analyses including previously reported HPA data of Malays, Chinese and Indians revealed details of the genetic relationships and ancestry of various sub-populations in Peninsular Malaysia. Furthermore, the comprehensive HPA allele frequency information from Peninsular Malaysia provided in this report has potential applications for future study of diseases, estimating risks associated with HPA alloimmunization and for developing an efficient HPA-typed donor recruitment strategy.


Subject(s)
Alleles , Antigens, Human Platelet/genetics , Gene Frequency , Genotype , Asian People , Female , Genotyping Techniques/methods , Humans , Malaysia , Male
3.
Int J Immunogenet ; 41(6): 472-9, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25367623

ABSTRACT

The KIR system shows variation at both gene content and allelic level across individual genome and populations. This variation reflects its role in immunity and has become a significant tool for population comparisons. In this study, we investigate KIR gene content in 120 unrelated individuals from the four Malay subethnic groups (Kelantan, Jawa, Banjar and Pattani Malays). Genotyping using commercial polymerase chain reaction-sequence-specific primer (PCR-SSP) kits revealed a total of 34 different KIR genotypes; 17 for Kelantan, 15 for Banjar, 14 for Jawa and 13 for Pattani Malays. Two new variants observed in Banjar Malays have not previously been reported. Genotype AA and haplotype A were the most common in Jawa (0.47 and 0.65, respectively), Banjar (0.37 and 0.52, respectively) and Pattani (0.40 and 0.60, respectively) Malays. In contrast, Kelantan Malays were observed to have slightly higher frequency (0.43) of genotype BB as compared with the others. Based on the KIR genes distribution, Jawa, Pattani and Banjar subethnic groups showed greater similarity and are discrete from Kelantan Malays. A principal component plot carried out using KIR gene carrier frequency shows that the four Malay subethnic groups are clustered together with other South-East Asian populations. Overall, our observation on prevalence of KIR gene content demonstrates genetic affinities between the four Malay subethnic groups and supports the common origins of the Austronesian-speaking people.


Subject(s)
Ethnicity/genetics , Genetic Variation , Receptors, KIR/genetics , Gene Frequency , Geography , Haplotypes/genetics , Humans , Malaysia , Principal Component Analysis
4.
Int J Immunogenet ; 40(6): 460-70, 2013 Dec.
Article in English | MEDLINE | ID: mdl-23870060

ABSTRACT

In recent years, with the application of genotyping technology, there has been a substantial increase in the number of reported blood group alleles. This survey was designed to evaluate new molecular blood group genotyping methods and compile reference blood group data sets for Polynesian and Maori subjects. Subsequent analyses of these results were used to calculate probability of random match, to trace Polynesian ancestry and migration patterns and to reveal past and present episodes of genetic admixture. Genomic DNA samples from Maori and Polynesian subjects were drawn from the Victoria University of Wellington DNA Bank and genotyped using combination of commercial PCR-SSP kits, hybridization SNP assay services or sequence-based typing. This survey also involves compilation of serological ABO and Rhesus blood group data from RakaiPaaka Iwi tribal members for comparison with those generated during our molecular blood group study. We observed perfect consistency between results obtained from all molecular methods for blood group genotyping. The A, O, DCcEe, DCCee, MNs, K-k+, Jk(a+b-), Jk(a+b+), Fy(a+b-), Fy(a+b+), Di(a+b-), Co(a+b-) and Do(a-b+) were predominant blood group phenotypes in both Polynesians and Maori. Overall, our survey data show only small differences in distributions of blood group phenotypes between Polynesian and Maori groups and their subgroups. These differences might be associated with selection, population history and gene flow from Europeans. In each case, we estimate that patients with certain blood groups have a very low probability of an exact phenotypic match, even if the patients were randomly transfused with blood from donors of their own ethnicity. The best way to avoid haemolytic transfusion reaction in such cases is to perform a pretransfusion cross-match and recruit increased numbers of donors with rare phenotype profiles. The conclusion of this study is that application of molecular method covering as many known variants as possible may help to improve the accuracy blood group genotyping and potentially conserve the routine requirements of transfusion centres.


Subject(s)
Blood Group Antigens/genetics , Genotyping Techniques/methods , Native Hawaiian or Other Pacific Islander/genetics , Transfusion Medicine/methods , Alleles , Blood Group Antigens/classification , Blood Grouping and Crossmatching/methods , DNA/genetics , Gene Frequency , Genotype , Humans , New Zealand , Phenotype , Phylogeny , Polymerase Chain Reaction/methods , Polymorphism, Single Nucleotide , Polynesia , Reproducibility of Results
5.
Transfus Med ; 23(5): 330-7, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23841727

ABSTRACT

BACKGROUND: Allele frequencies of human platelet antigens (HPA) reflect population history and possibility of platelet-specific alloimmunization. Here, we report on screening of variants at HPA loci for Polynesian and Maori subjects. OBJECTIVES: Our aims are to evaluate new HPA genotyping methods, compile and analyse new HPA datasets for these subjects, use HPA data for tracing ancestry, migration patterns, genetic admixture and its potential influence on health. MATERIALS AND METHODS: A total of 75 Maori and 25 Polynesian DNA samples were genotyped using commercial BAGene HPA-TYPE DNA-SSP kits, BLOODchip hybridization SNP assays and DNA sequence based typing. RESULTS: Genotyping was successful and cross validation of PCR-SSP and BLOODchip gave 100% agreement. Among the HPA loci tested, only six are dimorphic (HPA-1 to -3, -5, -6 and -15) and all others are monomorphic. The Polynesians and Maori have the 'a' allele form as the most common for all loci except HPA-15. CONCLUSIONS: The newly observed HPA data as well as principal coordinate analysis clearly indicate genetic contributions from both, Asia and Australasia in Maori and Polynesian populations together with recent admixture with Europeans. In addition, different prevalences of HPA alleles among Polynesian, Maori and European populations contribute towards different risk profiles for platelet-specific alloimmunization. This is the first report for these populations and our findings are of direct practical relevance for blood transfusion centres, the management of pregnancies, assessment of neonatal alloimmune thrombocytopenia and management of multi-transfused patients.


Subject(s)
Alleles , Antigens, Human Platelet/genetics , Genetic Loci , Native Hawaiian or Other Pacific Islander/genetics , Polymorphism, Single Nucleotide , Female , Genotyping Techniques , Humans , Male , New Zealand
6.
Hum Immunol ; 74(9): 1119-29, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23792058

ABSTRACT

Data from HLA typing studies have made significant contributions to genetic theories about the Austronesian diaspora and the health of descendant populations. To help further unravel pattern and process elements, we have typed HLA and MICA loci at high resolution in DNA samples from well defined groups of Maori and Polynesian individuals. Our results show a restricted set of HLA class I alleles compared with other well characterised populations. In contrast, the class II HLA-DRB1 locus seems to be diverse in Maori and Polynesians and both groups show high frequencies of HLA-DRB1(∗)04:03, -DRB1(∗)08:03, -DRB1(∗)09:01 and -DRB1(∗)12:01. Our survey also provides the first ever MICA datasets for Polynesians and reveal unusual distributions and associations with the HLA-B locus. Overall, our data provide further support for a hybrid origin for Maori and Polynesians. One novel feature of our study is the finding that the gene sequence of the HLA-B(∗)40:10 allele in Polynesians is a recombinant of HLA-B(∗)55:02 and -B(∗)40:01. HLA-B(∗)40:10 is in close association with HLA-C(∗)04:03, an allele identified as a hybrid of HLA-C(∗)04 and -C(∗)02. In this respect, our data resemble those reports on Amerindian tribes where inter-allele recombination has been a common means of generating diversity. However, we emphasize that Amerindian gene content per se is only a very minor element of the overall Polynesian genepool. The wider significance of HLA and MICA allele frequencies across the Pacific for modern day health is also discussed in terms of the frequency relative to reference populations of disease known to be associated with specific HLA and MICA markers. Thus, Polynesians and Maori are largely unaffected by "European autoimmune diseases" such as ankylosing spondylitis, uveitis and coeliacs disease, yet there are several Maori- and Polynesian-specific autoimmune diseases where the HLA and MICA associations are still to be determined.


Subject(s)
HLA Antigens/genetics , Histocompatibility Antigens Class I/genetics , Native Hawaiian or Other Pacific Islander/genetics , Alleles , Gene Frequency , Genotype , Health , Histocompatibility Testing , Humans , Linkage Disequilibrium , New Zealand , Polymorphism, Genetic , Polynesia , Recombination, Genetic
7.
Tissue Antigens ; 80(6): 509-22, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23137322

ABSTRACT

Human leukocyte antigens (HLA) are important genetic markers of tissue identity and accurately reflect ancestral history. The work reported in this paper provides a detailed description of HLA polymorphism in Polynesian and Maori individuals in relation to other populations. Our study concerns HLA classes I and II antigens in Polynesian (N = 36) and Maori (N = 114) subjects genotyped at two digit resolution by New Zealand Blood Service Laboratory in Auckland using polymerase chain reaction-sequence specific oligonucleotide and PCR-SSP technologies. We have also compared our data with those from other Austronesian-speaking Mongoloid and Papuan-speaking Australoid populations in order to test previously published account of the origin of Proto-Polynesians via gender-biassed gene flow between these two ancestral populations. We use principal coordinate analysis for this purpose, arguing this approach to be superior to tree-based methods, because of factors associated with population history and admixture. Our data are in general agreement with earlier work and reflect received wisdom on the dual origin of Proto-Polynesians. They also show the way in which the genetic make-up of Polynesian and Maori subjects is changing due to intermarriage with Europeans.


Subject(s)
Asian People/genetics , Ethnicity/genetics , HLA Antigens/genetics , Native Hawaiian or Other Pacific Islander/genetics , Female , Founder Effect , Gene Flow , Gene Frequency , Genes, MHC Class I , Genes, MHC Class II , Humans , Male , New Zealand , Polynesia
10.
Evolution ; 55(7): 1395-407, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11525463

ABSTRACT

New Zealand's isolation, its well-studied rapidly changing landscape, and its many examples of rampant speciation make it an excellent location for studying the process of genetic differentiation. Using 1520 base pairs of mitochondrial DNA from the cytochrome oxidase subunit I, ATPase subunits 6 and 8 and tRNA(Asp) genes, we detected two well-differentiated, parapatrically distributed clades within the widespread New Zealand cicada species Maoricicada campbelli that may prove to represent two species. The situation that we uncovered is unusual in that an ancient lineage with low genetic diversity is surrounded on three sides by two recently diverged lineages. Using a relaxed molecular clock model coupled with Bayesian statistics, we dated the earliest divergence within M. campbelli at 2.3 +/- 0.55 million years. Our data suggest that geological and climatological events of the late Pliocene divided a once-widespread species into northern and southern components and that near the middle of the Pleistocene the northern lineage began moving south eventually reaching the southern clade. The southern clade seems to have moved northward to only a limited extent. We discovered five potential zones of secondary contact through mountain passes that will be examined in future work. We predict that, as in North American periodical cicadas, contact between these highly differentiated lineages will exist but will not involve gene flow.


Subject(s)
DNA, Mitochondrial/genetics , Evolution, Molecular , Hemiptera/classification , Hemiptera/genetics , Phylogeny , Adenosine Triphosphatases/genetics , Animals , Base Sequence , Bayes Theorem , Electron Transport Complex IV/genetics , Environment , Genes, Insect/genetics , Genetic Variation , Geography , Haplotypes , Mitochondria/enzymology , Mitochondria/genetics , Molecular Sequence Data , New Zealand , RNA, Transfer/genetics
11.
Hum Mutat ; 17(4): 271-80, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11295824

ABSTRACT

An assessment of 28 pertinent binary genetic markers on the non-recombining portion of the Y chromosome (NRY) in New Zealand Maori and other relevant populations has revealed a diverse genetic paternal heritage of extant Maori. A maximum parsimony phylogeny was constructed in which nine of the 25 possible binary haplotypes were observed. Although approximately 40% of the samples have haplotypes of unequivocal European origin, an equivalent number of samples have a single binary haplotype that is also observed in Indonesia and New Guinea, indicative of common indigenous Melanesian ancestry. The balance of the lineages has either typical East Asian signatures or alternative compositions consistent with their affinity to Melanesia or New Guinea. Molecular analysis of mtDNA variation confirms the presence of a single predominant characteristic Southeast Asian (9-bp deletion in the Region V) lineage. The Y-chromosome results support a pattern of complex interrelationships between Southeast Asia, Melanesia, and Polynesia, in contrast to mtDNA and linguistic data, which uphold a rapid and homogeneous Austronesian expansion. The Y-chromosome data highlight a distinctive gender-modulated pattern of differential gene flow in the history of Polynesia.


Subject(s)
DNA, Mitochondrial/genetics , Ethnicity/genetics , Haplotypes/genetics , Phylogeny , White People/genetics , Y Chromosome/genetics , Chromatography, High Pressure Liquid , Female , Gene Frequency/genetics , Genetic Variation/genetics , Humans , Linguistics , Male , Microsatellite Repeats/genetics , Nucleic Acid Denaturation , Pacific Islands , Polymerase Chain Reaction , Polymorphism, Genetic/genetics
12.
J Arthroplasty ; 16(1): 87-91, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11172276

ABSTRACT

Sixty patients were prospectively assessed using the Western Ontario and McMaster Osteoarthritis Index (WOMAC) scale for osteoarthritis of the hip and the Short Form 36 (SF-36) general health status scale as well as the expectation WOMAC, which asked patients to estimate how they expected to feel 6 months after revision hip arthroplasty. There was a wide range of expectations, but we were unable to find any significant correlation between the patients' preoperative pain and stiffness levels and their expectations for pain and stiffness after revision hip arthroplasty. There was no significant correlation between the SF-36 scores and the patients' expectations. Our findings suggest that the expectations of patients awaiting revision hip arthroplasties are high and are not related closely to the level of preoperative disability.


Subject(s)
Arthroplasty, Replacement, Hip/psychology , Patient Satisfaction , Adult , Aged , Aged, 80 and over , Attitude to Health , Female , Humans , Male , Middle Aged , Osteoarthritis, Hip/psychology , Osteoarthritis, Hip/surgery , Prospective Studies , Surveys and Questionnaires
13.
Mol Biol Evol ; 18(2): 223-34, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11158381

ABSTRACT

The statistical testing of alternative phylogenetic trees is central to evaluating competing evolutionary hypotheses. Fleming proposed that the New Zealand cicada species Maoricicada iolanthe is the sister species to the major radiation of both low-altitude and montane Maoricicada species. However, using 1,520 bp of mitochondrial DNA sequence data from the cytochrome oxidase subunit I, tRNA aspartic acid, and the ATPase subunit 6 and 8 genes, we inferred that both M. iolanthe and another low-altitude species, Maoricicada campbelli, are nested within the montane Maoricicada radiation. Therefore, we examined the stability of the inferred phylogenetic placement of these two species using the newly developed Shimodaira-Hasegawa test (SH test) implemented in a maximum-likelihood framework. The SH test has two advantages over the more commonly used Kishino-Hasegawa (KH) and Templeton tests. First, the SH test simultaneously compares multiple topologies and corrects the corresponding P: values to accommodate the multiplicity of testing. Second, the SH test is correct when applied to a posteriori hypotheses, unlike the KH test, because it readjusts the expectation of the null hypothesis (that two trees are not different) accordingly. The comparison of P: values estimated under the assumptions of both the KH test and the SH test clearly demonstrate that the KH test has the potential to be misleading when the issue of comparing of a posteriori hypotheses is ignored and when multiple comparisons are not taken into account. The SH test, in combination with a variety of character-weighting schemes applied to our data, reveals a surprising amount of ambiguity in the phylogenetic placement of M. iolanthe and M. campbelli.


Subject(s)
Electron Transport Complex IV/genetics , Evolution, Molecular , Genetic Variation/genetics , Hemiptera/genetics , Invertebrates/classification , Mitochondria/enzymology , Animals , DNA Primers/chemistry , Hemiptera/classification , Hemiptera/enzymology , Invertebrates/enzymology , Invertebrates/genetics , New Zealand , Phylogeny , Polymerase Chain Reaction , Sequence Alignment , Species Specificity
14.
Syst Biol ; 50(1): 67-86, 2001 Feb.
Article in English | MEDLINE | ID: mdl-12116595

ABSTRACT

We have investigated the effects of different among-site rate variation models on the estimation of substitution model parameters, branch lengths, topology, and bootstrap proportions under minimum evolution (ME) and maximum likelihood (ML). Specifically, we examined equal rates, invariable sites, gamma-distributed rates, and site-specific rates (SSR) models, using mitochondrial DNA sequence data from three protein-coding genes and one tRNA gene from species of the New Zealand cicada genus Maoricicada. Estimates of topology were relatively insensitive to the substitution model used; however, estimates of bootstrap support, branch lengths, and R-matrices (underlying relative substitution rate matrix) were strongly influenced by the assumptions of the substitution model. We identified one situation where ME and ML tree building became inaccurate when implemented with an inappropriate among-site rate variation model. Despite the fact the SSR models often have a better fit to the data than do invariable sites and gamma rates models, SSR models have some serious weaknesses. First, SSR rate parameters are not comparable across data sets, unlike the proportion of invariable sites or the alpha shape parameter of the gamma distribution. Second, the extreme among-site rate variation within codon positions is problematic for SSR models, which explicitly assume rate homogeneity within each rate class. Third, the SSR models appear to give severe underestimates of R-matrices and branch lengths relative to invariable sites and gamma rates models in this example. We recommend performing phylogenetic analyses under a range of substitution models to test the effects of model assumptions not only on estimates of topology but also on estimates of branch length and nodal support.


Subject(s)
Models, Genetic , Phylogeny , Animals , Base Sequence , Biometry , DNA, Mitochondrial/genetics , Databases, Genetic , Evolution, Molecular , Genes, Insect , Genetic Variation , Hemiptera/classification , Hemiptera/genetics , Likelihood Functions , Molecular Sequence Data , Nucleic Acid Conformation , RNA, Transfer, Asp/chemistry , RNA, Transfer, Asp/genetics , Species Specificity
15.
Comp Biochem Physiol B Biochem Mol Biol ; 126(4): 455-76, 2000 Aug.
Article in English | MEDLINE | ID: mdl-11026658

ABSTRACT

Microsatellite DNA loci have recently been adopted for many biological applications. Comparative studies across a wide range of species has revealed many details of their mutational properties and evolutionary life cycles. Experience shows that a full understanding of these processes is essential to ensure the effective use of microsatellites as analytical tools. In this article, we review the controversies that have arisen as biologists have taken up this new technology and the emerging consensus that has resulted from their debates. We point to the need for comparative DNA sequencing studies to produce input data for a new generation of theoretical models of microsatellite behaviour. We conclude by presenting our own conceptual model, 'Snakes and Ladders', as an aid to theory development.


Subject(s)
Chromosomes/genetics , DNA/genetics , Microsatellite Repeats/genetics , Sequence Analysis, DNA/methods , Animals , Chromosomes/metabolism , DNA/classification , DNA/metabolism , Evolution, Molecular , Humans , Microsatellite Repeats/physiology , Mutation , Phylogeny
16.
J Urol ; 162(3 Pt 1): 699-701, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10458346

ABSTRACT

PURPOSE: We evaluated the use of intravesical potassium in the diagnosis of interstitial cystitis. MATERIALS AND METHODS: A blinded test assessment on 39 consecutive subjects attending our urology clinic for the evaluation of symptoms consistent with interstitial cystitis was performed. The pain response to intravesical potassium and water as a control was measured. The response rate was compared to the results of cystoscopy using standard outcome measures associated with diagnostic test assessment. RESULTS: The probability of having interstitial cystitis given a positive intravesical potassium test was 66%. This finding added no new useful information and would not be helpful with clinical decisions as the probability of having interstitial cystitis in this population was already 56% before the test. Similarly, if the test was negative then 46% or nearly half of the subjects were still likely to have interstitial cystitis. Therefore, a negative test would have no ability to rule out disease nor would it be useful in making clinical decisions about how to proceed with evaluation or therapy. Test characteristics were considered poor with a sensitivity of 69.5% and a specificity of 50%. Likelihood ratios (positive 1.39, negative 0.61) also indicated poor inclusion and exclusion capabilities. CONCLUSIONS: The general use of intravesical potassium as a diagnostic test for interstitial cystitis is not validated. The diagnosis of interstitial cystitis must depend on the clinical presentation and endoscopic findings based on National Institutes of Health criteria.


Subject(s)
Cystitis, Interstitial/diagnosis , Potassium Chloride , Administration, Intravesical , Adult , Female , Humans , Male , Potassium Chloride/administration & dosage , Single-Blind Method
19.
Biochim Biophys Acta ; 1386(1): 90-6, 1998 Jul 28.
Article in English | MEDLINE | ID: mdl-9675251

ABSTRACT

The albumin from an Atlantic salmonid, the brown trout (Salmo trutta), is 1730 Da higher in molecular mass than the albumin from a Pacific salmonid, the chinook salmon (Oncorhynchus tshawytscha), at 65230 Da. Digestion with neuraminidase revealed that purified brown trout albumin contained sialic acid while chinook salmon albumin did not. Concanavalin A-sepharose affinity chromatography was used to purify a glycopeptide from a total tryptic digest of brown trout albumin. The mass of this glycopeptide (3815 Da) was determined by mass spectrometry, and the sequence largely confirmed by N-terminal sequencing. The identified sequence of IAHCCNQSYSM-, contains an Asn-Gln-Ser glycosylation site and is identical to residues 475-485 derived from the cDNA of the albumin from the Atlantic salmon, the closest relative of the brown trout. Glycosylation of albumin is very unusual, and has not been identified in either reptilian or mammalian albumins. The finding of a glycoalbumin in salmonids, ancient members of the teleost fish subclass, coupled with evidence of albumin glycosylation in the oldest vertebrates, agnathans, as well as amphibians, suggests that albumin was originally a glycoprotein, but lost this modification sometime between the divergence of amphibians and reptiles.


Subject(s)
Glycoproteins/chemistry , Serum Albumin/chemistry , Trout , Animals , Evolution, Molecular , Glycopeptides/isolation & purification , Glycosylation , Protein Processing, Post-Translational , Salmon , Sequence Alignment , Sequence Analysis , Species Specificity , Vertebrates
20.
Comp Biochem Physiol B Biochem Mol Biol ; 117(2): 159-68, 1997 Jun.
Article in English | MEDLINE | ID: mdl-9226877

ABSTRACT

Vitellogenin (Vg), a major precursor to egg yolk proteins, was purified from plasma of an estradiol-treated female tuatara (Sphenodon punctatus) by MgCl2-EDTA precipitation and DEAE-cellulose chromatography. The amino acid composition of tuatara Vg is similar to that of other vertebtate Vgs and contains a large proportion of serine (13.7 mol/100 mol of total amino acid). The amino acid sequences of the N-terminus of mature Vg (33 residues) and of several trypsin- and CNBr-generated peptides were determined. Six peptide sequences obtained from tuatara Vg could be aligned with Vg sequences from other vertebrates. Reduced and non-reduced forms of tuatara Vg have the same apparent molecular mass (approximately 218 kDa) when resolved by SDS-PAGE, indicating that inter-chain disulfide bonds are not a feature of the molecule in this species. Western blot analysis with anti-tuatara Vg antiserum indicated that at least some epitopes are shared among Vgs of turtle, alligator and tuatara.


Subject(s)
Reptiles , Vitellogenins/blood , Vitellogenins/chemistry , Amino Acid Sequence , Amino Acids/analysis , Animals , Blotting, Western , Chemical Precipitation , Chromatography, DEAE-Cellulose , Cyanogen Bromide , Electrophoresis, Polyacrylamide Gel , Estradiol/pharmacology , Female , Magnesium Chloride , Molecular Sequence Data , Peptide Fragments/chemistry , Sequence Analysis , Species Specificity , Trypsin
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