Showing papers by "Rod Balhorn published in 1987"
TL;DR: The existence of sequence-specific nucleohistone and nucleoprotamine components within the human spermatozoon was demonstrated by cloning size-selected single-copy sequences and by using the derived clones as probes of nucleohistsone DNA and nucleobrotamine DNA.
Abstract: The DNA in human sperm chromatin is packaged into nucleoprotamine (approximately 85%) and nucleohistone (approximately 15%). Whether these two chromatin fractions are sequence-specific subsets of the spermatozoon genome is the question addressed in this report. Sequence-specific packaging would suggest distinct structural and functional roles for the nucleohistone and nucleoprotamine in late spermatogenesis or early development or both. After removal of histones with 0.65M NaCl, exposed DNA was cleaved with Bam HI restriction endonuclease and separated by centrifugation from insoluble nucleoprotamine. The DNA sequence distribution of nucleohistone DNA in the supernatant and nucleoprotamine DNA in the pellet was compared by cloning size-selected single-copy sequences and by using the derived clones as probes of nucleohistone DNA and nucleoprotamine DNA. Two clones derived from nucleohistone DNA preferentially hybridized to nucleohistone DNA, and two clones derived from nucleoprotamine DNA preferentially hybridized to nucleoprotamine DNA, which demonstrated the existence of sequence-specific nucleohistone and nucleoprotamine components within the human spermatozoon.
380 citations
TL;DR: All of the monoclonal antibodies except hupA are specific for human protamine, and suggest that each of the antibodies recognizes one of at least four distinct epitopes on protamine.
Abstract: Nine monoclonal antibodies to human protamine, hup1a, 1b, 1c, 1d, 1e, 2a, 2b, A, and B, have been isolated and partially characterized. Enzyme-linked immunoabsorption assay analyses with HPLC-separated human protamine 1 and protamine 2+3 mixture identified five of these antibodies as specific for human protamine 1, two antibodies specific for protamine 2+3 mixture and two monoclonal antibodies reactive with all three human protamines. These findings were confirmed by immunoblotting. None of the antibodies reacted with poly-arginine or somatic histone proteins. Additional analyses with bull, boar, and ram protamines indicated that all of the monoclonal antibodies except hupA are specific for human protamine. HupA reacted with protamines from all of the species tested. These studies suggest that each of the antibodies recognizes one of at least four distinct epitopes on protamine.
20 citations