Molecular Sizes of the Catalytic Units of Oxygen Evolution and the Reaction Center in Photosystem II of Spinach Thylakoids

Target analysis of the PS II reaction in spinach thylakoidsshowed that the respective molecular masses of the catalyticunits for oxygen evolution and the reaction center are about120 kDa and 250 kDa based on a kinetic separation of the tworeaction rates. The size of the oxygen-evolving enzyme agreedwith that determined for the PS II preparation from a thermophiliccyanobacterium by the same means [Nugent and Atkinson (1984)FEBS Lett. 170: 89]. Single hit-inactivation of oxygen evolutionand the PS II reaction center units indicates that each functionis driven by a structurally assembled unit. (Received August 6, 1984; Accepted December 17, 1984)     

GPAT2, a mitochondrial outer membrane protein,in piRNA biogenesis in germline stem cells

piRNA (PIWI-interacting RNA) is a germ cell–specific small RNA in which biogenesis PIWI (P-element wimpy testis) family proteins play crucial roles. MILI (mouse Piwi-like), one of the three mouse PIWI family members, is indispensable for piRNA production, DNA methylation of retrotransposons presumably through the piRNA, and spermatogenesis. The biogenesis of piRNA has been divided into primary and secondary processing pathways; in both of these MILI is involved in mice. To analyze the molecular function of MILI in piRNA biogenesis, we utilized germline stem (GS) cells, which are derived from testicular stem cells and possess a spermatogonial phenotype. We established MILI-null GS cell lines and their revertant, MILI-rescued GS cells, by introducing the Mili gene with Sendai virus vector. Comparison of wild-type, MILI-null, and MILI-rescued GS cells revealed that GS cells were quite useful for analyzing the molecular mechanisms of piRNA production, especially the primary processing pathway. We found that glycerol-3-phosphate acyltransferase 2 (GPAT2), a mitochondrial outer membrane protein for lysophosphatidic acid, bound to MILI using the cells and that gene knockdown of GPAT2 brought about impaired piRNA production in GS cells. GPAT2 is not only one of the MILI bound proteins but also a protein essential for primary piRNA biogenesis.     

Matrix-Embedded Osteocytes Regulate Mobilization of Hematopoietic Stem/Progenitor Cells

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Structural Basis for the Different Stability and Activity between the Cdk5 Complexes with p35 and p39 Activators

Cyclin-dependent kinase 5 (Cdk5) is a brain-specific membrane-bound protein kinase that is activated by binding to the p35 or p39 activator. Previous studies have focused on p35-Cdk5, and little is known regarding p39-Cdk5. The lack of functional understanding of p39-Cdk5 is due, in part, to the labile property of p39-Cdk5, which dissociates and loses kinase activity in nonionic detergent conditions. Here we investigated the structural basis for the instability of p39-Cdk5. p39 and p35 contain N-terminal p10 regions and C-terminal Cdk5 activation domains (AD). Although p35 and p39 show higher homology in the C-terminal AD than the N-terminal region, the difference in stability is derived from the C-terminal AD. Based on the crystal structures of the p25 (p35 C-terminal region including AD)-Cdk5 complex, we simulated the three-dimensional structure of the p39 AD-Cdk5 complex and found differences in the hydrogen bond network between Cdk5 and its activators. Three amino acids of p35, Asp-259, Asn-266, and Ser-270, which are involved in hydrogen bond formation with Cdk5, are changed to Gln, Gln, and Pro in p39. Because these three amino acids in p39 do not participate in hydrogen bond formation, we predicted that the number of hydrogen bonds between p39 and Cdk5 was reduced compared with p35 and Cdk5. Using substitution mutants, we experimentally validated that the difference in the hydrogen bond network contributes to the different properties between Cdk5 and its activators.     

Occurrence of Superoxide Dismutase in Bovine Milk

Acidic heteropolysaccharides, d-glucurono-d-xylo-d-mannans were isolated from the water- and alkaline extracts of the fruit body of Tremella fuciformis Berk. Similar polysaccharides were isolated from the growing culture of the haploid cells of two strains (T–19 and T–7) of T. fuciformis, when they were cultured in sucrose or glucose-yeast extract medium. The extracellular polysaccharides contain, d-glucuronic acid, d-xylose and d-mannose [molar ratios, 1.3: 1.0: 3.5 (T–7) and 0.8: 1.0: 2.1 (T–19)], and, in addition, small proportions of l-fucose and O-acetyl groups. Methylation and Smith degradation studies indicated that both fruit body and extracellular polysaccharides are built up of α-(1 → 3)-linked d-mannan backbone chain to which β-linked d-glucuronic acid and single or short chains of β-(1 → 2)-linked d-xylose residues are attached at the C–2 position. l-fucose residues in the extracellular polysaccharides may form the single branches. The structural features of these polysaccharides are discussed in comparison with the similar polysaccharides from other fungi.     

Continuous ATP Regeneration Using Immobilized Yeast Cells

Arthrobacter simplex was screened as an α-keto-δ-guanidinovalerate (ketoarginine) assimilating organism. A characteristic feature was its growth on ketoarginine as a carbon source; it began to grow after an extremely long lag. Its growth was stimulated by addition of 0.02% yeast extract to the medium.

The results indicated the transamination of arginine-α-ketoglutarate (α-KGA) and the hydrolyzing reaction of ketoarginine into α-keto-δ-aminovalerate and urea. Two intermediates, ketoarginine and α-keto-δ-aminovalerate, were isolated and identified by various procedures. Coupling of the two reactions was demonstrated in cell-free extracts of arginine-grown cells; ketoarginine formed from arginine by transamination with α-KGA was hydrolyzed directly to α-keto-δ-aminovalerate and urea. The metabolic routes of arginine in microorganisms were discussed.