In human spermatozoa, 15-20% of histones are retained in the nucleus to coexist with protamines. Hypothetically, nucleohistone regions of sperm chromatin mark DNA sequences for distinctive processing during fertilization and early embryogenesis. The structural organization and molecular composition of nucleohistones in human spermatozoa is poorly studied. Here, we isolate and characterize fractions of sperm chromatin that are solubilized by endogenous and micrococcal nucleases. Chromatin isolated by either nuclease have a nucleosomal organization with the periodicity of approximately 195 bp (endogenous nuclease digest) and approximately 189 bp (micrococcal nuclease digest), which is similar to that of somatic cells. A distinct feature of sperm nucleohistone is its specific compact supra-nucleosomal organization that was demonstrated by two-dimensional electrophoresis and by atomic force microscopy. The latter technique showed compacted fiber arrays composed of globular particles with the prevailing diameter of approximately 16 nm. A rough estimation indicates that histones may cover continuous stretches of >50 kbp of sperm DNA. This initial characterization of sperm chromatin solubilized by nucleases is important for our understanding of the bipartite structural organization of the paternal genome.