cis-Terpin Hydrate Metabolism by a Brevibacterium: Patterns of Enzyme Induction, and Accumulation of α-Terpineol in Growth Media

A brevibacterium, strain TH-4, previously isolated by aerobic enrichment on the monocyclic monoterpenoid cis-terpin hydrate as a sole carbon and energy source, was found to grow on alpha-terpineol and on a number of common sugars and organic acids. Oxidation of these terpenoids was shown to occur via an induced enzyme system, as measured manometrically by oxygen uptake and prevention of protein synthesis with chloramphenicol or puromycin. Oxidation of terpin hydrate by cell suspensions appeared to be coincidentally induced by growth on alpha-terpineol, and oxidation of alpha-terpineol similarly appeared to be induced by growth on terpin hydrate. Culture fluids in which the TH-4 organism was grown at the expense of cis-terpin hydrate were found to contain (-)-alpha-terpineol in combined butanol-ether extracts. The isolated compound was shown to be chromatographically and spectrophotometrically identical to an authentic sample of alpha-terpineol. The stereospecificity of an enzymatic dehydration of terpin hydrate to alpha-terpineol is considered.     

Some observations on the breeding, age structure, dispersion and habitat of populations of Macropus robustus and Macropus antilopinus (Marsupialia)

Differences in breeding, population structure, dispersion and habitat are described between various species and subspecies of wallaroo (Macropus robustus robustus; M. r. cervinus; M. r. alligatoris; M. r. erubescens; Macropus antilopinus; Macropus bernardus).
Pouch young of Macropus robustus erubescens in western New South Wales were born throughout the year, while in both M. r. alligatoris and M. antilopinus then Northern Territory, most pouch young found were born during March and April.
In the populations of wallaroos in western New South Wales and the Northern Territory where there had been no systematic shooting of wallaroos for many years, 11|X% of the animals were immature. In the New England district of New South Wales where regular shooting occurs, 46|X% of the animals were immature.
The habitat of M. r. alligatoris of the Northern Territory was very similar to that of M. erubescens in inland Australia, amongst rocky hills and gullies. M. antilopinus , which is sympatric with M. r. alligatoris in the Northern Territory, also occurred in the rocky hills, but it was also found in open savannah woodland in flat and gently undulating country.
M. r. erubescens and M. r. alligatoris were almost always seen alone or in pairs, while M. antilopinus often formed larger groups.     

Identification of a cellular protein substrate phosphorylated by the avian sarcoma virus-transforming gene product

The avian sarcoma virus-transforming gene product (pp60src) appears potentially able to mediate cell transformation via phosphorylation since it is tightly associated with a protein kinase activity. We have searched for and have been able to identify a normal cellular protein that appears to be a substrate of pp60src. The phosphorylation of this protein (34K) in transformation-specific in ASV-transformed cells of both avian and mammalian origin. Moreover, the 34K polypeptide serves as a substrate for the pp60src phosphotransferase activity in vitro and is phosphorylated at a site identical to the major site of phosphorylation in vivo. These data suggest that upon transformation the 34,000-dalton protein is phosphorylated directly as a result of pp60src activity.     

Cell junctions in early embryos of squid (Loligo pealei)

Summary Squid embryos examined by freeze-fracture and thin-section electron microscopy exhibit identifiable gap junctions during mid-cleavage stages (stages 7–8), and junctional complexes composed of adherent appositions, elaborate septate junctions and gap junctions at slightly later stages (stages 12–13). During germinal layer establishment (stages 12–13) cytoplasmic bridges frequently link the embryonic cells. The presence of gap junctions in cleavagestage embryos provides the morphological substrate for a demonstrated pathway of direct cell-cell communication that is modifiable by experimental treatments and may be physiologically regulatable. The existence of septate junctions and adherent contacts at later stages suggests that some functional specialization, perhaps the establishment of a strongly joined framework of cells at the surface of the embryo, accompanies the formation of germinal layers.     

The Active Site of a Carbohydrate Esterase Displays Divergent Catalytic and Noncatalytic Binding Functions

Multifunctional proteins, which play a critical role in many biological processes, have typically evolved through the recruitment of different domains that have the required functional diversity. Thus the different activities displayed by these proteins are mediated by spatially distinct domains, consistent with the specific chemical requirements of each activity. Indeed, current evolutionary theory argues that the colocalization of diverse activities within an enzyme is likely to be a rare event, because it would compromise the existing activity of the protein. In contrast to this view, a potential example of multifunctional recruitment into a single protein domain is provided by CtCel5C-CE2, which contains an N-terminal module that displays cellulase activity and a C-terminal module, CtCE2, which exhibits a noncatalytic cellulose-binding function but also shares sequence identity with the CE2 family of esterases. Here we show that, unlike other CE2 members, the CtCE2 domain displays divergent catalytic esterase and noncatalytic carbohydrate binding functions. Intriguingly, these diverse activities are housed within the same site on the protein. Thus, a critical component of the active site of CtCE2, the catalytic Ser-His dyad, in harness with inserted aromatic residues, confers noncatalytic binding to cellulose whilst the active site of the domain retains its esterase activity. CtCE2 catalyses deacetylation of noncellulosic plant structural polysaccharides to deprotect these substrates for attack by other enzymes. Yet it also acts as a cellulose-binding domain, which promotes the activity of the appended cellulase on recalcitrant substrates. The CE2 family encapsulates the requirement for multiple activities by biocatalysts that attack challenging macromolecular substrates, including the grafting of a second, powerful and discrete noncatalytic binding functionality into the active site of an enzyme. This article provides a rare example of “gene sharing,” where the introduction of a second functionality into the active site of an enzyme does not compromise the original activity of the biocatalyst.     

Crystal Structure of the 2-Oxoglutarate- and Fe(II)-Dependent Lysyl Hydroxylase JMJD6

Lysyl and prolyl hydroxylations are well-known post-translational modifications to animal and plant proteins with extracellular roles. More recent work has indicated that the hydroxylation of intracellular animal proteins may be common. JMJD6 catalyses the iron- and 2-oxoglutarate-dependent hydroxylation of lysyl residues in arginine-serine-rich domains of RNA-splicing-related proteins. We report crystallographic studies on the catalytic domain of JMJD6 in complex with Ni(II) substituting for Fe(II). Together with mutational studies, the structural data suggest how JMJD6 binds its lysyl residues such that it can catalyse C-5 hydroxylation rather than N^?-demethylation, as for analogous enzymes.