Association Study of AMP-Activated Protein Kinase Subunit Genes in Polycystic Ovary Syndrome

Total Page:16

File Type:pdf, Size:1020Kb

Association Study of AMP-Activated Protein Kinase Subunit Genes in Polycystic Ovary Syndrome European Journal of Endocrinology (2009) 161 405–409 ISSN 0804-4643 CLINICAL STUDY Association study of AMP-activated protein kinase subunit genes in polycystic ovary syndrome Kari Sproul1,2, Michelle R Jones3, Ricardo Azziz1,2,4 and Mark O Goodarzi1,3,4,5 1Department of Obstetrics and Gynecology, Cedars-Sinai Medical Center, Los Angeles, California 90048, USA, 2Department of Obstetrics and Gynecology, the David Geffen School of Medicine at UCLA, Los Angeles, California 90095, USA, 3Division of Endocrinology, Diabetes and Metabolism, Department of Medicine, Cedars-Sinai Medical Center, 8700 Beverly Boulevard, Room B-131, Los Angeles, California 90048, USA, 4Department of Medicine, the David Geffen School of Medicine at UCLA, Los Angeles, California 90095, USA and 5Medical Genetics Institute, Cedars-Sinai Medical Center, Los Angeles, California 90048, USA (Correspondence should be addressed to M O Goodarzi at Division of Endocrinology, Diabetes and Metabolism, Department of Medicine, Cedars-Sinai Medical Center; Email: [email protected]) Abstract Objective: To examine the genes for AMP-activated protein kinase (AMPK) subunits a2(PRKAA2) and g3(PRKAG3) as candidates for polycystic ovary syndrome (PCOS) and its component traits. Design and methods: A total of 287 white PCOS women were recruited from the reproductive endocrinology clinic at the University of Alabama at Birmingham and 187 white control subjects were recruited from the surrounding community. Seven PRKAA2 single nucleotide polymorphisms (SNPs) and four PRKAG3 SNPs were genotyped in PCOS cases and controls. Genotyping and association analysis were performed at Cedars-Sinai Medical Center. Results: Nominal associations of PRKAA2 variants with insulin-related traits and the PRKAG3 Pro71Ala variant with PCOS were not statistically significant after multiple testing correction. Among PCOS patients, there were no associations between variants in AMPK subunit genes and androgenic or reproductive traits. Conclusions: Variants in genes for AMPKa2 and AMPKg3 were not associated with PCOS or its component traits. Our evidence does not demonstrate that AMPK is a major genetic risk factor for PCOS. European Journal of Endocrinology 161 405–409 Introduction PCOS ovarian theca cells, suggesting that metformin stimulation of AMPK in the ovary itself may promote Polycystic ovary syndrome (PCOS) is a common complex ovulation (7). We therefore examined the genes for genetic disorder, affecting w6–8% of reproductive age AMPKa2(PRKAA2) and AMPKg3(PRKAG3)as women (1, 2). PCOS is characterized by hyperandro- candidate genes for PCOS and its component traits. genism, ovulatory dysfunction, polycystic ovarian morphology, and frequently insulin resistance (IR). AMP-activated protein kinase (AMPK) is a cellular energy sensor, acting to maintain ATP levels in the cell Subjects and methods by sensing the AMP/ATP ratio and then phosphorylat- ing metabolic enzymes and regulating gene expression Subjects and phenotyping with the net result of activating processes that generate A total of 287 consecutive unrelated white patients with ATP and inhibiting processes that consume ATP. AMPK PCOS, aged 13–47 years, and 187 healthy unrelated influences food intake, fatty acid and glucose uptake in white control women, aged 14–60, were recruited from skeletal and cardiac muscle, insulin secretion, and the Birmingham, AL, USA area. PCOS cases were hepatic fatty acid synthesis and gluconeogenesis (3). recruited from the reproductive endocrine practice of Consequently, AMPK subunit genes have been one of the investigators (RA) at the University of examined as candidates for type 2 diabetes suscep- Alabama at Birmingham (UAB). All subjects were tibility, with conflicting results (4, 5). non-Hispanic Caucasians. Participation in research As PCOS is a metabolic disorder associated with an studies was offered to patients meeting inclusion increased risk of diabetes, AMPK may play a role in this criteria (premenopausal, not pregnant, on no hormonal association. Metformin, commonly prescribed for PCOS therapy for at least 3 months, and meeting diagnostic patients, activates AMPK activity in skeletal muscle (6). criteria for PCOS). PCOS was diagnosed following the In addition, AMPK activation appears to be decreased in 1990 NIH consensus criteria (8). q 2009 European Society of Endocrinology DOI: 10.1530/EJE-09-0245 Online version via www.eje-online.org Downloaded from Bioscientifica.com at 09/27/2021 12:40:30AM via free access 406 K Sproul and others EUROPEAN JOURNAL OF ENDOCRINOLOGY (2009) 161 Controls were healthy women with a history of Statistical analysis regular menstrual cycles and no family history of 2 hirsutism. These women had no evidence of hirsutism, Unpaired t-tests and c tests were used to compare acne or alopecia, or endocrine dysfunction and had not clinical characteristics between women with and taken hormonal therapy (including oral contraceptives) without PCOS; quantitative trait values were log- or for at least 3 months prior to testing. Controls were square root-transformed as appropriate to reduce non- recruited by word of mouth and advertisements in normality. the Birmingham, Alabama area, through a call for Association with the presence/absence of PCOS and ‘healthy women’ without detailing further the nature qualitative traits was evaluated using logistic of the studies. regression, adjusting for age and body mass index Subjects underwent a physical examination and (BMI) by including both as independent variables. serum androgen measurement, as per a previously Association with quantitative traits was evaluated described protocol (1). Fasting glucose and insulin were using analysis of covariance, again adjusting for age also obtained in a subset of the cohort (w70%). This and BMI. Recognizing that insulin secretion responds to subset did not differ demographically or hormonally ambient IR, HOMA-%B associations were also calcu- from the study subjects overall. The computer-based lated with HOMA-IR as a covariate in addition to age homeostasis model assessment (HOMA, www.dtu.ox.ac. and BMI. Quantitative trait associations were conducted in the PCOS and control groups separately. To handle uk/homa) utilizes fasting glucose and insulin to ! calculate indices of IR (HOMA-IR) and insulin secretion multiple testing, significance was taken as P 0.006, (HOMA-%B) (9). For the insulin-related traits only, six considering that we analyzed two genes against four PCOS subjects with diabetes were excluded because the families of traits (PCOS diagnosis, androgens, metabolic hyperglycemia of diabetes may induce secondary traits and reproductive endpoints), yielding a Bonfer- changes in insulin-related traits that reduce their utility roni correction factor of 8 (i.e. eight independent for genetic analysis. In addition, in PCOS cases only, we comparisons). Association analyses excluding 26 evaluated the following reproductive traits: severe post-menopausal subjects from the control group oligomenorrhea (defined as O3monthsbetween yielded the same results as analyses conducted in the menses), infertility (yes/no), and parity (yes/no). entire cohort; therefore, the latter results are reported. Clinical characteristics of the subjects are given in supplementary Table 1, which can be viewed online at http://www.eje-online.org/supplemental/. All subjects gave written informed consent, and the Results study was performed according to the guidelines of the Institutional Review Boards of UAB and Cedars- PRKAA2 Sinai Medical Center, Los Angeles, CA, USA. We genotyped seven SNPs spanning the PRKAA2 gene (Fig. 1A). SNP frequencies are shown in Table 1. All Genetic analysis markers were in Hardy–Weinberg equilibrium. Linkage disequilibrium among these markers (D0) ranged from We selected six PRKAA2 single nucleotide polymorph- 0.45 to 1.0 (average pairwise D0 of 0.90). The r2 values isms (SNPs; rs11206887, rs2143749, rs2746349, ranged from 0 to 0.98 (average 0.23). rs12749128, rs857155, and rs3738568) using geno- Among the women with PCOS, carriers of the minor type data of the Caucasian population of the HapMap alleles of SNPs rs2143749, rs2746349, or rs857155 database (release 24, phase II) (10). These SNPs capture had higher fasting insulin levels than noncarriers 28 out of 35 (80%) of the Caucasian HapMap SNP (PZ0.044, 0.021, and 0.027 respectively). Carriers of alleles at r2O0.8. An additional PRKAA2 SNP the minor allele of SNP rs2143749 also had increased (rs2051040) was selected because it was associated HOMA-%B (PZ0.030), which was not significant when with type 2 diabetes in a Japanese cohort (4). Four HOMA-IR was also included as a covariate (PZ0.26). validated PRKAG3 SNPs (rs6436094, rs16859382, Lastly, PCOS women who were carriers of the minor rs692243, and rs650898) were genotyped, selected allele of SNP rs2746349 had increased HOMA-IR based on frequency O5% because only two SNPs compared with noncarriers (PZ0.039). Trait values in HapMap (rs6436094, rs692243) are polymorphic for PCOS women are shown in the electronic supple- in Caucasians. SNP rs692243 is a nonsynonymous mental Table 3, which can be viewed online at http:// variant (proline to alanine at codon 71). The eleven www.eje-online.org/supplemental/.Noneofthese SNPs were genotyped in the 474 subjects using PRKAA2 associations were significant after considering the 50-exonuclease assay (TaqMan MGB, Applied multiple testing. We did not find significant associations Biosystems, Foster City, CA, USA). Supplementary between PRKAA2 variantsandPCOSitself,BMI,
Recommended publications
  • Gene Symbol Gene Description ACVR1B Activin a Receptor, Type IB
    Table S1. Kinase clones included in human kinase cDNA library for yeast two-hybrid screening Gene Symbol Gene Description ACVR1B activin A receptor, type IB ADCK2 aarF domain containing kinase 2 ADCK4 aarF domain containing kinase 4 AGK multiple substrate lipid kinase;MULK AK1 adenylate kinase 1 AK3 adenylate kinase 3 like 1 AK3L1 adenylate kinase 3 ALDH18A1 aldehyde dehydrogenase 18 family, member A1;ALDH18A1 ALK anaplastic lymphoma kinase (Ki-1) ALPK1 alpha-kinase 1 ALPK2 alpha-kinase 2 AMHR2 anti-Mullerian hormone receptor, type II ARAF v-raf murine sarcoma 3611 viral oncogene homolog 1 ARSG arylsulfatase G;ARSG AURKB aurora kinase B AURKC aurora kinase C BCKDK branched chain alpha-ketoacid dehydrogenase kinase BMPR1A bone morphogenetic protein receptor, type IA BMPR2 bone morphogenetic protein receptor, type II (serine/threonine kinase) BRAF v-raf murine sarcoma viral oncogene homolog B1 BRD3 bromodomain containing 3 BRD4 bromodomain containing 4 BTK Bruton agammaglobulinemia tyrosine kinase BUB1 BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast) BUB1B BUB1 budding uninhibited by benzimidazoles 1 homolog beta (yeast) C9orf98 chromosome 9 open reading frame 98;C9orf98 CABC1 chaperone, ABC1 activity of bc1 complex like (S. pombe) CALM1 calmodulin 1 (phosphorylase kinase, delta) CALM2 calmodulin 2 (phosphorylase kinase, delta) CALM3 calmodulin 3 (phosphorylase kinase, delta) CAMK1 calcium/calmodulin-dependent protein kinase I CAMK2A calcium/calmodulin-dependent protein kinase (CaM kinase) II alpha CAMK2B calcium/calmodulin-dependent
    [Show full text]
  • Brief Genetics Report Haplotype Structures and Large
    Brief Genetics Report Haplotype Structures and Large-Scale Association Testing of the 5؅ AMP-Activated Protein Kinase Genes PRKAA2, PRKAB1, and PRKAB2 With Type 2 Diabetes Maria W. Sun,1,2 Jennifer Y. Lee,1,2 Paul I.W. de Bakker,1,2,3 Noe¨l P. Burtt,2 Peter Almgren,4 Lennart Råstam,5 Tiinamaija Tuomi,6 Daniel Gaudet,7 Mark J. Daly,2,8 Joel N. Hirschhorn,2,3,9 David Altshuler,1,2,3,8,10 Leif Groop,4,6 and Jose C. Florez1,2,8,10 AMP-activated protein kinase (AMPK) is a key molecular plasma glucose, or insulin sensitivity. Several nominal asso- regulator of cellular metabolism, and its activity is induced ciations of variants in PRKAA2 and PRKAB1 with BMI appear by both metformin and thiazolidinedione antidiabetic med- to be consistent with statistical noise. Diabetes 55:849–855, ications. It has therefore been proposed both as a putative 2006 agent in the pathophysiology of type 2 diabetes and as a valid target for therapeutic intervention. Thus, the genes that encode the various AMPK subunits are intriguing ype 2 diabetes arises from the complex interplay candidates for the inherited basis of type 2 diabetes. We therefore set out to test for the association of common of various pathophysiologic mechanisms involv- variants in the genes that encode three selected AMPK ing peripheral insulin resistance and relative subunits with type 2 diabetes and related phenotypes. Of Tinsulin insufficiency. The final expression of the the seven genes that encode AMPK isoforms, we initially diabetic phenotype is strongly influenced by inheritance; chose PRKAA2, PRKAB1, and PRKAB2 because of their however, with the exception of rare monogenic forms of higher prior probability of association with type 2 diabetes, diabetes, common type 2 diabetes is thought to have a based on previous reports of genetic linkage, functional polygenic architecture (1).
    [Show full text]
  • Transcriptomic Characterization of Fibrolamellar Hepatocellular
    Transcriptomic characterization of fibrolamellar PNAS PLUS hepatocellular carcinoma Elana P. Simona, Catherine A. Freijeb, Benjamin A. Farbera,c, Gadi Lalazara, David G. Darcya,c, Joshua N. Honeymana,c, Rachel Chiaroni-Clarkea, Brian D. Dilld, Henrik Molinad, Umesh K. Bhanote, Michael P. La Quagliac, Brad R. Rosenbergb,f, and Sanford M. Simona,1 aLaboratory of Cellular Biophysics, The Rockefeller University, New York, NY 10065; bPresidential Fellows Laboratory, The Rockefeller University, New York, NY 10065; cDivision of Pediatric Surgery, Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, NY 10065; dProteomics Resource Center, The Rockefeller University, New York, NY 10065; ePathology Core Facility, Memorial Sloan-Kettering Cancer Center, New York, NY 10065; and fJohn C. Whitehead Presidential Fellows Program, The Rockefeller University, New York, NY 10065 Edited by Susan S. Taylor, University of California, San Diego, La Jolla, CA, and approved September 22, 2015 (received for review December 29, 2014) Fibrolamellar hepatocellular carcinoma (FLHCC) tumors all carry a exon of DNAJB1 and all but the first exon of PRKACA. This deletion of ∼400 kb in chromosome 19, resulting in a fusion of the produced a chimeric RNA transcript and a translated chimeric genes for the heat shock protein, DNAJ (Hsp40) homolog, subfam- protein that retains the full catalytic activity of wild-type PKA. ily B, member 1, DNAJB1, and the catalytic subunit of protein ki- This chimeric protein was found in 15 of 15 FLHCC patients nase A, PRKACA. The resulting chimeric transcript produces a (21) in the absence of any other recurrent mutations in the DNA fusion protein that retains kinase activity.
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • 32-3099: PRKAB1 Recombinant Protein Description
    9853 Pacific Heights Blvd. Suite D. San Diego, CA 92121, USA Tel: 858-263-4982 Email: [email protected] 32-3099: PRKAB1 Recombinant Protein Alternative Name : AMPK,HAMPKb,5'-AMP-activated protein kinase subunit beta-1,AMPK subunit beta-1,AMPKb,PRKAB1. Description Source : E.coli. PRKAB1 Human Recombinant produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 293 amino acids (1-270 a.a.) and having a molecular mass of 32.8 kDa. The PRKAB1 is fused to a 23 amino acid His Tag at N- Terminus and purified by proprietary chromatographic techniques. 5'-AMP-activated protein kinase subunit beta-1 (PRKAB1) hinders protein, carbohydrate and lipid biosynthesis, in addition to cell growth and proliferation. AMPK is a heterotrimer comprised of an alpha catalytic subunit, and non-catalytic beta and gamma subunits. AMPK acts via direct phosphorylation of metabolic enzymes, and longer-term effects by phosphorylation of transcription regulators. PRKAB1 is a regulator of cellular polarity by remodeling the actin cytoskeleton; most likely by indirectly activating myosin. Beta non-catalytic subunit acts as a scaffold on which the AMPK complex compiles, through its C-terminus that joins alpha (PRKAA1 or PRKAA2) and gamma subunits (PRKAG1, PRKAG2 or PRKAG3). Product Info Amount : 5 µg Purification : Greater than 85% as determined by SDS-PAGE. The PRKAB1 protein solution (0.5mg/ml) contains 20mM Tris-HCl buffer (pH 8.0), 0.15M NaCl, Content : 10% glycerol and 1mM DTT. Store at 4°C if entire vial will be used within 2-4 weeks. Store, frozen at -20°C for longer periods of Storage condition : time.
    [Show full text]
  • Age, DNA Methylation and the Malignant Potential of the Serrated Neoplasia Pathway Lochlan John Fennell B
    Age, DNA Methylation and the Malignant Potential of the Serrated Neoplasia Pathway Lochlan John Fennell B. Biomed Sci A thesis submitted for the degree of Doctor of Philosophy at The University of Queensland in 2020 Faculty of Medicine ORC ID: 0000-0003-3214-3527 1 Abstract Colorectal cancer is the third most common cancer in Australia and is responsible for the death of over four thousand Australians each year. There are two overarching molecular pathways leading to colorectal cancer. The conventional pathway, which is responsible for ~75% of colorectal cancer diagnoses, occurs in a step-wise manner and is the consequence of a series of genetic alterations including mutations of tumour suppressor genes and gross chromosomal abnormalities. This pathway has been extensively studied over the past three decades. The serrated neoplasia pathway is responsible for the remaining colorectal cancers. This pathway is triggered by oncogenic BRAF mutation and these cancers accumulate epigenetic alterations while progressing to invasive cancer. DNA methylation is important in serrated neoplasia, however the extent and role of DNA methylation on the initiation and progression of serrated lesions is not clear. DNA methylation accumulates in tissues with age, and advanced serrated lesions and cancers occur almost exclusively in elderly patients. How this methylation affects serrated lesions is unknown. In this thesis I set out to address three key research questions related to DNA methylation, age and serrated colorectal neoplasia. First, what is the extent of DNA methylation in colorectal cancers?; Second, Does age-related hypermethylation, and namely that occurring at the loci encoding tumour suppressor genes, increase the risk of serrated colorectal neoplasia?; and if true, how can we reconcile this with the existence of early onset serrated colorectal cancer? In the first chapter of this thesis, I examine the DNA methylation and transcriptional architecture of 216 colorectal cancer samples collected consecutively at the Royal Brisbane and Women’s hospital.
    [Show full text]
  • AMPK Beta 1 (PRKAB1) Mouse Monoclonal Antibody [Clone ID: OTI3D10] Product Data
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for TA813116S AMPK beta 1 (PRKAB1) Mouse Monoclonal Antibody [Clone ID: OTI3D10] Product data: Product Type: Primary Antibodies Clone Name: OTI3D10 Applications: WB Recommended Dilution: WB 1:500 Reactivity: Human, Mouse, Rat Host: Mouse Isotype: IgG1 Clonality: Monoclonal Immunogen: Human recombinant protein fragment corresponding to amino acids 2-270 of human PRKAB1 (NP_006244) produced in E.coli. Formulation: PBS (PH 7.3) containing 1% BSA, 50% glycerol and 0.02% sodium azide. Concentration: 1 mg/ml Purification: Purified from mouse ascites fluids or tissue culture supernatant by affinity chromatography (protein A/G) Conjugation: Unconjugated Storage: Store at -20°C as received. Stability: Stable for 12 months from date of receipt. Predicted Protein Size: 30.2 kDa Gene Name: protein kinase AMP-activated non-catalytic subunit beta 1 Database Link: NP_006244 Entrez Gene 19079 MouseEntrez Gene 83803 RatEntrez Gene 5564 Human Q9Y478 This product is to be used for laboratory only. Not for diagnostic or therapeutic use. View online » ©2021 OriGene Technologies, Inc., 9620 Medical Center Drive, Ste 200, Rockville, MD 20850, US 1 / 2 AMPK beta 1 (PRKAB1) Mouse Monoclonal Antibody [Clone ID: OTI3D10] – TA813116S Background: Non-catalytic subunit of AMP-activated protein kinase (AMPK), an energy sensor protein kinase that plays a key role in regulating cellular energy metabolism. In response to reduction of intracellular ATP levels, AMPK activates energy-producing pathways and inhibits energy- consuming processes: inhibits protein, carbohydrate and lipid biosynthesis, as well as cell growth and proliferation.
    [Show full text]
  • The Study of Copy Number Variations in the Regions of PRKAB2 and PPM1K Among Congenital Heart Defects Patients
    The study of copy number variations in the regions of PRKAB2 and PPM1K among congenital heart defects patients Han-Quan Dong1 Yue-Xin Du2 1. Department of Pneumology, Tianjin Children’s Hospital, Tianjin, 300074–China 2. Department of Child Healthcare, Tianjin Municipal Women and Children health care center, Tianjin, 300070, China http://dx.doi.org/10.1590/1806-9282.65.6.786 SUMMARY OBJECTIVE: This study was to assess the genetic association of copy number variations in two genes (PRKAB2 and PPM1K) located in two regions (tetralogy of Fallot and ventricular septal defect) in a Chinese Han population. METHODS: A total of 200 congenital heart disease patients (100 tetralogy of Fallot patients and 100 ventricular septal defect patients) and 100 congenital heart defect-free controls were recruited, and quantitative real-time PCR analysis was used to replicate the asso- ciation of two copy number variations with congenital heart defects in a Chinese Han population. RESULTS: One deletion at PRKAB2 and one duplication at PPM1K were found in two of the tetralogy of Fallot patients, respectively; while all these regions were duplicated in both ventricular septal defect patients and in the 100 congenital heart defects-free controls. CONCLUSIONS: We replicated the copy number variations at the disease-candidate genes of PRKAB2 and PPM1K with tetralogy of Fallot in a Chinese Han population, and in patients with ventricular septal defect mutations in these two genes were not found. These results indicate the same molecular population genetics exist in these two genes with different ethnicity. This shows that these two genes are possibly specific pf tetralogy of Fallot candidates.
    [Show full text]
  • Polymorphisms in AKT3, FIGF, PRKAG3, and TGF-Beta Genes Are
    Research Note Polymorphisms in AKT3, FIGF, PRKAG3, and TGF-β genes are associated with myofiber characteristics in chickens Sirui Chen ,1 Jianyong An ,1 Ling Lian , Lujiang Qu , Jiangxia Zheng , Guiyun Xu , and Ning Yang 2 National Engineering Laboratory for Animal Breeding and MOA Key Laboratory of Animal Genetics and Breeding, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China ABSTRACT Muscle characteristics such as myofiber for each bird. Six SNP with a very low minor allele fre- diameter, density, and total number are important quency (<1%) were excluded for further analysis. The traits in broiler breeding and production. In the pres- remaining 13 SNP were used for the association study ent study, 19 SNP of 13 major genes, which are located with muscle characteristics. The results showed that in the vicinity of quantitative trait loci affecting breast SNP in TGF-β1/2/3 had significant effects on myofiber muscle weight, including INS, IGF2, PIK3C2A, AKT3, diameter. A SNP in PRKAG3 had a significant effect PRKAB2, PRKAG3, VEGFA, RPS6KA2/3, FIGF, and on myofiber density (P < 0.05). A C > G mutation in TGF-β1/2/3, were chosen to be genotyped by high- FIGF was strongly associated with total fiber number throughput matrix-assisted laser desorption/ionization (P < 0.05). Additionally, birds with the GG genotype time-of-flight mass spectrometry in a broiler popula- of the C > G mutation in AKT3 had significantly larger tion. One hundred twenty birds were slaughtered at 6 myofiber numbers (P < 0.05) than birds with the CC or wk of age.
    [Show full text]
  • AMPK Beta 2 (PRKAB2) Mouse Monoclonal Antibody [Clone ID: OTI4H4] Product Data
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for TA808333 AMPK beta 2 (PRKAB2) Mouse Monoclonal Antibody [Clone ID: OTI4H4] Product data: Product Type: Primary Antibodies Clone Name: OTI4H4 Applications: WB Recommended Dilution: WB 1:2000 Reactivity: Human, Mouse, Rat Host: Mouse Isotype: IgG1 Clonality: Monoclonal Immunogen: Full length human recombinant protein of human PRKAB2 (NP_005390) produced in E.coli. Formulation: PBS (PH 7.3) containing 1% BSA, 50% glycerol and 0.02% sodium azide. Concentration: 1 mg/ml Purification: Purified from mouse ascites fluids or tissue culture supernatant by affinity chromatography (protein A/G) Conjugation: Unconjugated Storage: Store at -20°C as received. Stability: Stable for 12 months from date of receipt. Predicted Protein Size: 30.1 kDa Gene Name: protein kinase AMP-activated non-catalytic subunit beta 2 Database Link: NP_005390 Entrez Gene 64562 RatEntrez Gene 5565 Human O43741 This product is to be used for laboratory only. Not for diagnostic or therapeutic use. View online » ©2021 OriGene Technologies, Inc., 9620 Medical Center Drive, Ste 200, Rockville, MD 20850, US 1 / 2 AMPK beta 2 (PRKAB2) Mouse Monoclonal Antibody [Clone ID: OTI4H4] – TA808333 Background: The protein encoded by this gene is a regulatory subunit of the AMP-activated protein kinase (AMPK). AMPK is a heterotrimer consisting of an alpha catalytic subunit, and non-catalytic beta and gamma subunits. AMPK is an important energy-sensing enzyme that monitors cellular energy status. In response to cellular metabolic stresses, AMPK is activated, and thus phosphorylates and inactivates acetyl-CoA carboxylase (ACC) and beta-hydroxy beta- methylglutaryl-CoA reductase (HMGCR), key enzymes involved in regulating de novo biosynthesis of fatty acid and cholesterol.
    [Show full text]
  • Active AMPK (A1/B2/G3)
    Catalog # Aliquot Size P83-10G-05 5 µg P83-10G-10 10 µg AMPK (A1/B2/G3), Active Full-length recombinant protein expressed in Sf9 cells Catalog # P83-10G Lot # Y1103 -1 Product Description Specific Activity Recombinant full-length human AMPK (combination of A1/B2/G3 subunits) was expressed by baculovirus in Sf9 480,000 insect cells using the N-terminal GST and C-terminal His tags. The gene accession numbers for the three subunits 360,000 (A1/B2/G3) are NM_006251 , NM_005399 , and NM_017431 . 240,000 Gene Aliases 120,000 Activity (cpm) Activity Subunit A1: PRKAA1, MGC33776, MGC57364 0 Subunit B2: PRKAB2, MGC61468 0 50 100 150 200 Subunit G3: PRKAG3 Protein (ng) Formulation The specific activity of AMPK was determined to be 400 nmol /min/mg as per activity assay protocol. Recombinant protein stored in 50mM Tris-HCl, pH 7.5, 150mM NaCl, 10mM glutathione, 0.1mM EDTA, 0.25mM Purity DTT, 0.1mM PMSF, 25% glycerol. Storage and Stability Store product at –70 oC. For optimal storage, aliquot target into smaller quantities after centrifugation and The purity of AMPK was determined store at recommended temperature. For most favorable to be >80% by densitometry, performance, avoid repeated handling and multiple approx. MW ~92kDa (A1), ~62kDa (B2), and ~108kDa (G3). freeze/thaw cycles. Scientific Background AMP-activated protein kinase (AMPK) exhibits a key role as a master regulator of cellular energy homeostasis (1). AMPK exists as a heterotrimeric complex composed of a AMPK (A1/B2/G3), Active catalytic α subunit and regulatory β and γ subunits. Full-length recombinant protein expressed in Sf9 cells Binding of AMP to the γ subunit allosterically activates the complex.
    [Show full text]
  • Development and Validation of a Protein-Based Risk Score for Cardiovascular Outcomes Among Patients with Stable Coronary Heart Disease
    Supplementary Online Content Ganz P, Heidecker B, Hveem K, et al. Development and validation of a protein-based risk score for cardiovascular outcomes among patients with stable coronary heart disease. JAMA. doi: 10.1001/jama.2016.5951 eTable 1. List of 1130 Proteins Measured by Somalogic’s Modified Aptamer-Based Proteomic Assay eTable 2. Coefficients for Weibull Recalibration Model Applied to 9-Protein Model eFigure 1. Median Protein Levels in Derivation and Validation Cohort eTable 3. Coefficients for the Recalibration Model Applied to Refit Framingham eFigure 2. Calibration Plots for the Refit Framingham Model eTable 4. List of 200 Proteins Associated With the Risk of MI, Stroke, Heart Failure, and Death eFigure 3. Hazard Ratios of Lasso Selected Proteins for Primary End Point of MI, Stroke, Heart Failure, and Death eFigure 4. 9-Protein Prognostic Model Hazard Ratios Adjusted for Framingham Variables eFigure 5. 9-Protein Risk Scores by Event Type This supplementary material has been provided by the authors to give readers additional information about their work. Downloaded From: https://jamanetwork.com/ on 10/02/2021 Supplemental Material Table of Contents 1 Study Design and Data Processing ......................................................................................................... 3 2 Table of 1130 Proteins Measured .......................................................................................................... 4 3 Variable Selection and Statistical Modeling ........................................................................................
    [Show full text]