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Isolation, Fractionation, and Analysis of Nonhistone Chromosomal Proteins
Published in Lubomir S. Hnilica, Chromosomal Nonhistone Proteins, 2018
Leokadia Klyszejko-Stefanowicz, Lubomir S. Hnilica
In the procedure of MacGillivray and co-workers,162,426,427 chromatin was dissociated in 2 M NaCl-5 M urea-1 mM sodium phosphate buffer, pH 6.8, with addition of 2 mM TrisCl to protect the proteins against cyanate degradation,155,156 and 0.1 mM PMSF as protease inhibitor.32 Its chromatography on a HAP column provided five distinct fractions which were eluted with increasing concentrations of sodium phosphate (50 to 200 mM) and 2 M GdnCl (HAP4 and HAP5). The HAP1 fraction (over 70% of total chromatin protein) not retained in the loading (i.e., dissociating) buffer, contained histones and no nucleic acid material. However, contamination of this fraction with up to 15% of NHCP was suggested by [3H]-tryptophan labeling. The HAP2 fraction, eluted with 50 mM sodium phosphate and containing about 2% RNA, represented just over 40% of the total NHCP (about 12% of total chromatin protein). These proteins were much more phosphorylated than those of the HAP1 fraction. Proteins of the HAP3 fraction, eluted with 200 mM sodium phosphate, were two to three times more phosphorylated than the HAP2 proteins. HAP3 contained relatively large amounts of RNA (up to 50% of the total protein) and, occasionally, small amounts of DNA.
Structural vaccinology of malaria transmission-blocking vaccines
Published in Expert Review of Vaccines, 2021
PfHAP2 (PlasmoDB: PF3D7_1014200) is a member of the HAP2/GCS1 family of proteins that are class II fusion proteins with a cysteine-rich extracellular region [33]. The HAP2/GCS1 family consists of conserved male-specific proteins expressed on the male gametocyte and microgamete, and are essential for mediating membrane fusion during fertilization [19]. Disruption of PbHAP2 in P. berghei affects the ability of male gametes to fuse with female gametes and interrupts the essential process of fertilization required for subsequent parasite stages in the mosquito [34,96]. The conserved fusion loop (cd loop) is comprised of 174 to 205 amino acid residues in PbHAP2 and from 178 to 207 amino acid residues in PfHAP2, and has been demonstrated to elicit transmission-blocking immunity [19]. This suggests that the cd loop may be essential for gamete fusion and is a potential TBV target [19]. PbHAP2 and PfHAP2 have shown to elicit functional antibodies in rodents [33,97]. Future structural definition of this essential protein family is expected to drive the development of PfHAP2 as a TBV candidate.
Filling the patient–provider knowledge gap: a patient advocate to address asthma care and self-management barriers
Published in Journal of Asthma, 2019
Anna M. Localio, Heather L. Black, Hami Park, Luzmercy Perez, Grace Ndicu, Heather Klusaritz, Marisa Rogers, Xiaoyan Han, Andrea J. Apter
This qualitative study was conducted within the context of a larger clinical randomized-controlled trial: the ongoing Helping Asthma Patients 2 (HAP2) Study [11]. HAP2 investigates whether a PA intervention is associated with improved asthma control and other asthma-related health outcomes for adults with moderate to severe asthma. Patient participants were randomized 1:1 to the PA intervention or usual care. The intervention was administered over a 6-month period, and data collection was performed with all participants five times over the course of 1 year. Written informed consent was obtained from all participants. The University of Pennsylvania Institutional Review Board granted ethical clearance to the protocol. The study is registered at ClinicalTrials.gov (NCT01972308).
Transmission-Blocking Vaccines: Harnessing Herd Immunity for Malaria Elimination
Published in Expert Review of Vaccines, 2021
Additional TBV candidates will enter the clinic in future, starting with Pfs48/45. The female gamete surface antigen Pfs47 mediates evasion of the mosquito immune system [46] by interacting with a specific midgut receptor [47] and is thought to thereby determine mosquito receptivity to parasite variants carrying specific Pfs47 alleles. In addition to its role in parasite–mosquito interactions, Pfs47 also induces transmission-blocking antibodies in animals; the specificity of these antibodies has been mapped to a central 52 amino acid (aa) region [48]. The pre-fertilization antigen HAP2/GCS1 (Hapless 2/Generative Cell-Specific 1), which is involved in male fertility, has also been shown to induce transmission-blocking antibodies in preclinical studies [49–52].