Determination of a common genetic variant of luteinizing hormone using DNA hybridization and immunoassays

Nilsson, Christel; Jiang, Min; Pettersson, Kim; Iitiä, Antti; Mäkelä, Minna; Simonsen, Henrik Toft; Easteal, Simon; Herrera, René J.; Huhtaniemi, Ilpo

doi:10.1046/j.1365-2265.1998.00532.x

Cited by 49 publications

(25 citation statements)

References 29 publications

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“…19q13.3) (Pettersson et al ., 1994). The two SNPs coding for V‐LH (A‐>G, rs1800447; A‐>G, rs34349826) are located in the LHB exon 2 and exhibit complete linkage disequilibrium (LD) (Nilsson et al ., 1998). V‐LH seems to be a universally common variant with the average minor allele frequency among Europeans 8% (range 7.34–11.8%; Huhtaniemi et al ., 2010).…”

Section: Introductionmentioning

confidence: 99%

‘Carriers of V‐LH among 1593 Baltic men have significantly higher serum LH’

et al. 2015

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SummaryLuteinizing hormone (LH) is a pituitary heterodimeric glycoprotein essential in male and female reproduction. Its functional polymorphic variant (V‐LH) is determined by two missense mutations (rs1800447, A/G, Trp8Arg; rs34349826, A/G, Ile15Thr) in the LH β‐subunit encoding gene (LHB; 19q13.3; 1111 bp; 3 exons). Among women, V‐LH has been associated with higher circulating LH and reduced fertility, but the knowledge of its effect on male reproductive parameters has been inconclusive. The objective of this study was to assess the effect of V‐LH on hormonal, seminal and testicular parameters in the Baltic young men cohort (n = 986; age: 20.1 ± 2.1 years) and Estonian idiopathic infertility patients (n = 607; 35.1 ± 5.9 years). V‐LH was detected by genotyping of the underlying DNA polymorphisms using PCR‐RFLP combined with resequencing of a random subset of subjects. Genetic associations were tested using linear regression under additive model and results were combined in meta‐analysis. No significant difference was detected between young men and infertility patients for the V‐LH allele frequency (11.0 vs. 9.3%, respectively). V‐LH was associated with higher serum LH in both, the young men cohort (p = 0.022, allelic effect = 0.26 IU/L) and the idiopathic infertility group (p = 0.008, effect = 0.59 IU/L). In meta‐analysis, the statistical significance was enhanced (p = 0.0007, resistant to Bonferroni correction for multiple testing; effect = 0.33 IU/L). The detected significant association of V‐LH with increased serum LH remained unchanged after additional adjustment for the SNPs previously demonstrated to affect LH levels (FSHB ‐211G/T, FSHR Asn680Ser, FSHR ‐29A/G). Additionally, a suggestive trend for association with reduced testicular volume was observed among young men, and with lower serum FSH among infertility patients. The V‐LH carrier status did not affect sperm parameters and other circulating reproductive hormones. For the first time, we show a conclusive contribution of V‐LH to the natural variance in male serum LH levels. Its downstream clinical consequences are still to be learned.

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Section: Introductionmentioning

confidence: 99%

‘Carriers of V‐LH among 1593 Baltic men have significantly higher serum LH’

et al. 2015

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“…As a result, in the TaqI region the non-restriction percents were 87. 5 The non-restriction percent of homozygous showed values of 100 or 0 neighborhood, and almost heterozygous showed about 50%. However, in the case of four samples (18, 21, 23 and 24) which were a heterozygous sample, the non-restriction percents of the BsmI region polymorphism were 83, 79, 72 and 75, respectively.…”

Section: Application To Clinical Samplesmentioning

confidence: 99%

“…In recent years, easy and rapid detection methods of PCR products using hybridization 4,5 and immunochromatography (IC) 6,7 have been developed, though there are several problems: for instance, the hybridization method requires detection step by a probe and a denature step; also IC is not sensitive, dose not have a high throughput, and is not suitable for the analysis of gene polymorphism.…”

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“…However, there are a few reports related to gene analysis. 5,15 In this study, we describe the development of a simultaneous detection method for the measuring the duplex PCR-RFLP products. Thus, the BsmI recognition site and the TaqI recognition site on the VDR gene were analyzed by timeresolved fluorescence immunoassay (TR-FIA) with two different lanthanide ion-chelate labels.…”

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Simultaneous Analysis of Two-Site Gene Polymorphisms Using Time-Resolved Fluorescence Immunoassay

et al. 2000

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“…Following the failure to detect serum LH in some patients by two-site immunoassays that utilised highly specific monoclonal antibody pairs (Pettersson et al 1991), mutations altering the antigenic structure of the LH -subunit were identified. Of particular interest is a genetic variant of LH (vLH) that arises from two single point mutations in the LH gene that each alters the primary amino acid sequence: Trp8<Arg (TGG<CGG) and Ile15<Thr (ATC<ACC) (Furui et al 1994, Pettersson et al 1994, Nilsson et al 1998). An extra glycosylation consensus sequence (Asn-X-Thr/Ser) is thereby introduced into the LH -subunit by the second mutation which, it may be speculated, might permit the attachment of an N-linked oligosaccharide moiety at Asn13.…”

Section: Introductionmentioning

confidence: 99%

Identification of post-translational modifications resulting from LHbeta polymorphisms by matrix-assisted laser desorption time-of-flight mass spectrometric analysis of pituitary LHbeta core fragment

Jacoby

Kicman

Iles³

2003

Journal of Molecular Endocrinology

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Metabolism of the human chorionic gonadotrophin (hCG)-and LHβ-subunits (hCGβ, LHβ) terminates with the urinary excretion of core fragment (hCGβcf, LHβcf) molecules that retain antigenic shape and constituent N-linked carbohydrate moieties. We have previously demonstrated the resolved mass spectra of hCGβcf, from which the carbohydrate moieties present at two N-linked glycosylation sites were identified. LHβcf was subjected to the same mass spectrometric analysis. As LHβ shares 82% homology with hCGβ but possesses only one glycosylation consensus site a simpler spectral fingerprint of LHβcf glycoforms was expected. LHβcf was reduced with dithiothreitol and analysed by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. Glycoforms were predicted by subtracting the peptide mass from the m/z values of the observed peaks and then sequentially subtracting the masses of the monosaccharide residues of hCGβ N-linked carbohydrates reported in the literature. The mass spectra of LHβcf revealed a broad single peak ranging from m/z 8700 to 10 700. Following reduction, this peak was replaced by a set of partially resolved peaks between m/z 4130 and 5205 corresponding to glycosylated forms of the peptide LHβ6-40. A peak at m/z 4252·2 corresponded to the non-glycosylated peptide LHβ55-93. Remaining peaks indicated that the pooled sample comprised a wide set of glycoforms, contained LHβcf with two N-linked carbohydrate moieties and indicated evidence of further glycosylation due to amino acid substitution in polymorphic variants. This is evidence that a single nucleotide polymorphism alters the post-translational modification of a protein and hence its structural phenotype.

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Determination of a common genetic variant of luteinizing hormone using DNA hybridization and immunoassays

Abstract: The immunoassay technique and the hybridization assay can be used as alternatives to determine the LH status. A great variation in carrier frequency of the V-LH beta allele is observed in different populations.

Cited by 49 publications

References 29 publications

‘Carriers of V‐LH among 1593 Baltic men have significantly higher serum LH’

‘Carriers of V‐LH among 1593 Baltic men have significantly higher serum LH’

Simultaneous Analysis of Two-Site Gene Polymorphisms Using Time-Resolved Fluorescence Immunoassay

Identification of post-translational modifications resulting from LHbeta polymorphisms by matrix-assisted laser desorption time-of-flight mass spectrometric analysis of pituitary LHbeta core fragment

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