15N NMR relaxation studies of calcium-loaded parvalbumin show tight dynamics compared to those of other EF-hand proteins.

Dynamics of the rat alpha-parvalbumin calcium-loaded form have been determined by measurement of 15N nuclear relaxation using proton-detected heteronuclear NMR spectroscopy. The relaxation data were analyzed using spectral density functions and the Lipari-Szabo formalism. The major dynamic features for the rat alpha-parvalbumin calcium-loaded form are (1) the extreme rigidity of the helix-loop-helix EF-hand motifs and the linker segment connecting them, (2) the N and C termini of the protein being restricted in their mobility, (3) a conformational exchange occurring at the kink of helix D, and (4) the residue at relative position 2 in the Ca2+-binding sites having an enhanced mobility. Comparison of the Ca2+-binding EF-hand domains of alpha-parvalbumin-Ca2+, calbindin-Ca2+, and calmodulin-Ca2+ shows that parvalbumin is probably the most rigid of the EF-hand proteins. It also illustrates the dynamical properties which are conserved in the EF-hand domains from different members of this superfamily: (1) a tendency toward higher mobility of NH vectors at relative position 2 in the Ca2+-binding loop, (2) a restricted mobility for the other residues in the binding loop, and (3) an overall rigidity for the helices of EF-hand motifs. The differences in mobility between parvalbumin and the two EF-hand proteins occur mainly at the linker connecting the pair of EF hands and also at the C terminus of the last helix. In parvalbumin-Ca2+, these two regions are characterized by a pronounced rigidity compared to the corresponding more mobile regions in calbindin-Ca2+ and calmodulin-Ca2+.

Kinetics of amide proton exchange in parvalbumin studied by 1H 2-D NMR. A comparison of the calcium and magnesium loaded forms.

The amide proton exchange rates have been measured for the pike parvalbumin loaded either with calcium (PaCa2) or with magnesium (PaMg2) by using 2-D total correlation spectroscopy experiments. The differences in the exchange rates observed between these two species were unexpected when compared with the small conformational changes induced in parvalbumin by the Ca/Mg exchange. With the calcium-loaded protein (PaCa2), a significant difference was observed for the amide proton exchange rates of residues located in the N-terminal domain AB in contrast to the slower exchange rates that were observed in the CD and EF domains. Such a difference does not exist for PaMg2, where faster exchange rates are observed over all the sequence. Since amide proton exchange rates are the signature of the solvent's accessibility in proteins, we interpreted our results in terms of difference of the equilibria between 'closed-states' and 'opened-states' for individual amide protons of the protein when calcium was replaced by magnesium. The CD and EF domains, and to a lesser extent the AB domain, would be more rigid when the protein was loaded with calcium ions. For the magnesium-loaded parvalbumin (PaMg2) the faster exchange rates we observed could be rationalized by a more flexible structure than in the case of the PaCa2.

Characterization of Micrococcaceae strains isolated from the human urogenital tract by the conventional scheme and a micromethod.

Two hundred and twelve Micrococcaceae isolates were obtained from 82 men with nongonococcal urethritis, 24 women with vaginitis, and 54 girls with vulvovaginitis. Identification and biotyping of these strains were carried out by using the simplified scheme of Kloos and Schleifer (W. E. Kloos and K. H. Schleifer, J. Clin. Microbiol. 1:82-88, 1975) and the commercially available API Staph test (DMS Staph Trac). Staphylococcus epidermidis occurred in about half of these isolates. There was no statistical difference between the urethral and vaginal specimens, except for S. haemolyticus found primarily in males and for S. simulans and S. aureus found primarily in girls between the ages of 1 and 12 years. S. saprophyticus, a major cause of urinary tract infections in young women, was never isolated from the vagina, suggesting the probability of another reservoir.

[Stomatococcus mucilaginosus: cultural and biochemical properties of 100 strains isolated from the ORL area]. / Stomatococcus mucilaginosus: caractères culturaux et biochimiques de 100 souches isolées de la sphère ORL.

A study of the growth characteristics and 80 biochemical tests using API System micromethods (API-50CH, API-20E, API-Zym) was carried out on 100 strains of Stomatococcus mucilaginosus isolated from the human oral cavity. Seventy-seven other enzymatic tests from the API system (AP-arylamidases, osidases and esterases) were also performed on 10 of these strains. The results of these tests were compared to those of 24 type strains of Micrococcaceae and Aerococcus viridans. A simple identification scheme for S. mucilaginosus is proposed.