An Eco R1 polymorphism of a human platelet factor 4 (PF4) gene

Blood, 1987

We report the isolation of a platelet factor 4 (PF4) cDNA clone from a lambda gt11 expression cDNA library which was derived from a human erythroleukemic (HEL) cell line. The sequence of the DNA insert includes the 3′-untranslated region, the entire amino acid coding region for the mature PF4 protein, and a 5′ region containing coding information for an additional 18 amino acids. In addition, supplemental genomic DNA sequencing shows that the full-length leader sequence is 30 amino acids long plus an initial methionine and codes for a hydrophobic signal-like sequence which is probably involved in transmembrane transport. A single species mRNA of approximately 800 nucleotides was detected on blots of HEL cell poly(A) + RNA using a labeled PF4 cDNA probe. The human PF4 leader sequence shares some DNA, but no amino acid, homology with the 15 amino acids at the N-terminus of mature bovine PF4, suggesting rapid divergence in this region of PF4 between these two species. Sequence comparis...

Blood, 1990

Platelet factor 4 (PF4) is a 70 amino acid heparin-binding protein released from the a-granules of activated platelets. Its exact biologic function is not known, although PF4 is a member of a multigene family involved in chemotaxis, coagulation, inflammation, and cell growth. We previously cloned the cDNA for human PF4 from a human erythroleukemic (HEL) cell expression library. We now report the isolation and sequence determination of the gene for human PF4. This gene contains three exons and spans approximately 1 ,OOO basepairs (bp). Concurrently, we have cloned a highly homologous gene that we have called PF4, , , . We show that PF4 and PF4, , are non-allelic genes: the human PF4 gene is encoded on a 10 kilobasepairs (kb) EcoRl fragment, and its DNA sequence agrees with protein and cDNA data for PF4. while PF4, , is encoded in a polymorphic 3 or 5 kb EcoRl fragment. Compared with PF4, this gene has 14% DNA and 38% amino acid divergence in the signal peptide region, and 2.6% DNA and 4.3% amino acid divergence in the coding region of the mature protein.

1993

factor 4 Selection of an HEL-derived cell line expressing high levels of platelet http://bloodjournal.hematologylibrary.org/site/misc/rights.xhtml#repub_requests

Thrombosis Research, 1980

The localization of platelet factor 4 (PF4) in the megakaryocyte has been determined by indirect immunofluorescent studies using monospecific rabbit antibody to human PFt,. Also, specific staining has demonstrated that platelets in peripheral blood contain PFk antigen. Absorption studies established that the staining was specific for PFq. Using this technique, it has been demonstrated that PF, is also localized in the megakaryocytes in bone marrow. The existence of PFI, antigen in the megakaryocytes suggests that PF,+ may be synthesized in the megakaryocytes.

American Journal of Human Genetics, 1992

Protein 4.2 (P4.2), one of the major components of the red-blood-cell membrane, is located on the interior surface, where it binds with high affinity to the cytoplasmic domain of band 3. Individuals whose red blood cells are deficient in P4.2 have osmotically fragile, abnormally shaped cells and moderate hemolytic anemia. cDNA clones from both the 5â² and the 3â² coding

Journal of Cell Biology, 1983

The site of synthesis of platelet-specific proteins remains to be established. With the use of short-term megakaryocyte-enriched cultures, direct evidence was obtained to show that megakaryocytes synthesize the platelet-specific protein, platelet factor 4. A megakaryocyteenriched fraction of rabbit bone marrow for culture was obtained by centrifugal elutriation and cultured with [3H]leucine. Newly synthesized aH-platelet factor 4 was sought by copurification with added carrier rabbit platelet factor 4, using heparin agarose affinity chromatography and immunoprecipitation with specific goat anti-rabbit platelet factor 4 antisera. SDS PAGE of the washed immunoprecipitates demonstrated a [aH]leucine-containing peak which migrated identically with purified homogeneous rabbit platelet factor 4. A second, slightly larger molecular-weight protein was identified in the gels also, suggesting that rabbit platelet factor 4 may be synthesized as a larger molecular-weight precursor in rabbit megakaryocytes. These results provide direct evidence that the platelet-specific protein, platelet factor 4, is synthesized in rabbit megakaryocytes before it is packaged into a-granules for release in circulating platelets.

Transfusion, 1996

Background: Platelet-specific antibodies may be involved in refractoriness to platelet transfusions, disorders such as neonatal alloimmune thrombocytopenia, and posttransfusion purpura. Genotyping for the major human platelet antigen (HPA) systems HPA-1 through HPA-5 is of considerable help in establishing the diagnoses of these diseases. Study Design and Methods: A new genotyping method is described. Alleles of all five systems are amplified in a multiplex polymerase chain reaction. Subsequently, aliquots of the amplification products are thermocycled in the presence of a pair of allele-specific oligonucleotide probes and a heat-stable ligase. After heat denaturation, the probes hybridize adjacent to complementary sequences of the amplification product. In a perfect match, the two probes become covalently joined. Detection of the ligation product is performed with an enzyme-linked immunosorbent assay. Results: Complete concordance of genotypes between the ligation-based typing and established genotyping methods was determined in 54 Austrian (HPA-1,-2,-3, and-5) and 56 Japanese (HPA-4) individuals. Ligation-based genotyping of HPA-1 polymorphism using platelet-derived RNA as starting material gave concordant results in all 15 cases tested. Conclusion: Multiplex polymerase chain reaction in combination with ligation-based typing allows fast typing of large numbers of platelet donors and screening for critical antigens in pregnant women. Abbreviations: ASRA = allele-specific restriction enzyme analysis; ELISA =enzymelinked immunosorbent assay; GP = glycoprotein; HPA = human platelet antigen; MAIPA = monoclonal antibody-specific immobilization of platelet antigens (assay); NAIT = neonatal alloimmune thrombocytopenia; OD = optical density; PCR = polymerase chain reaction; PTP = posttransfusion purpura; SSP = sequence-specific primers. PLATELET-SPECIFIC ANTIBODIES may be involved in patients with neonatal alloimmune thrombocytopenia (NAIT), refractoriness to platelet transfusions, or posttransfusion purpura (PTP). The five human platelet antigen (HPA) systems, designated HPA-1 through HPA-5, have initially been defined by the use of antibodies obtained from immunized patients. Alleles of these systems are distributed differently among several populations studied. Molecular analyses have revealed that the polymorphism can be traced to single nucleotide substitutions in platelet glycoprotein (GP) genes.',* As to the nomenclature, several designations have been proposed; in this article, we will use the HPA nomenclature introduced by von dem Borne et al.3

Seminars in Thrombosis and Hemostasis, 2019

Recent advances in genetic analysis are bringing huge benefits to patients with rare genetic disorders, including those with inherited disorders of platelet number and function. Modern clinical hematological practice now has a range of genetic techniques available to enable the precision diagnosis of inherited platelet disorders. There are some features of this disparate group of inherited disorders that present specific challenges to establishing an accurate genetic diagnosis. This review aims to introduce the techniques that are relevant for the genetic diagnosis of inherited platelet disorders and will discuss the key considerations necessary for their application to the clinic.

Transfusion, 1996

Background: Thre~nine'~~/methionine'~~ dimorphism in platelet glycoprotein (GP) Iba defines the human platelet antigen (HPA)Q system that has been implicated in refractoriness to HLA-matched platelet transfusion and in neonatal immune thrombocytopenic purpura.

JURNAL PENELITIAN DAN KARYA ILMIAH LEMBAGA PENELITIAN UNIVERSITAS TRISAKTI, 2018

The need for coal as fuel in the industrial field still plays an important role, as evidenced by the increasing demand for coal with certain quality to date. The purpose of this research are: To know the availability of coal in the research area, to know the direction of coal deployment and its thickness, to know the prospect of coal in the research area. The expected target is to know the prospect of coal in the research area. By using survey method observation and observation field then correlation and analysis based on USGS method.According to Setyana, 2005 regional geology of research areas into the Ketungau Basin, so that the sedimentary rocks in it have the same deposition history on the Basin. The formation of a coal carrier is a formation of mites. Coal outcrops are very rare in research areas with average stance and slope N 45oE/12o. From the results of the correlation of several test wells and the analysis of outcrop data, the coal in the study area has a thickness between...