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At least 1,297 records · Page 72Linked to original sources
PubMed1999

C-terminal domains of human translation termination factors eRF1 and eRF3 mediate their in vivo interaction.

At the termination step of protein synthesis, hydrolysis of the peptidyl-tRNA is jointly catalysed at the ribosome by the termination codon and the polypeptide release factor (eRF1 in eukaryotes). eRF1 forms in vivo and in vitro a stable complex with release factor eRF3, an eRF1-dependent and ribosome-dependent GTPase. The role of the eRF1-eRF3 complex in translation remains unclear. We have undertaken a systematic analysis of the interactions between the human eRF1 and eRF3 employing a yeast two-hybrid assay. We show that the N-terminal parts of eRF1 (positions 1-280) and of eRF3 (positions 1477) are either not involved or non-essential for binding. Two regions in each factor are critical for mutual binding: positions 478-530 and 628-637 of eRF3 and positions 281-305 and 411-415 of eRF1. The GTP binding domain of eRF3 is not involved in complex formation with eRF1. The GILRY pentamer (positions 411-415) conserved in eukaryotes and archaebacteria is critical for eRF1's ability to stimulate eRF3 GTPase. The human eRF1 lacking 22 C-terminal amino acids remains active as a release factor and promotes an eRF3 GTPase activity whereas C-terminally truncated eRF3 is inactive as a GTPase.

Amino Acid Sequence↗
PubMed1999

Sensitivity of single-equivalent trunk extensor muscle models to anatomical and functional assumptions.

Single-equivalent muscle models are often used to estimate loads on the lumbosacral joint after net extension moments have been calculated by means of inverse dynamics. These models usually ignore the effects trunk flexion has on the extensor lever arm. In addition, no systematic analysis of the sensitivity to the anatomical and functional assumptions made in these models is available. In the present study a series of single-equivalent models incorporating trunk flexion dependence was derived from a detailed description of the trunk musculature. Each model was based on different anatomical and functional assumptions. The differences of estimates of compression and shear forces on the lumbosacral disc during a lifting movement resulting from these models were analysed. The results show that these load estimates heavily depend on assumptions regarding anthropometry, lumbar curvature and coactivity of abdominal muscles and only moderately on assumptions regarding force sharing between extensor muscles. Fairly simple single-equivalent models with the net moment and thorax orientation as input can be used to predict lumbosacral compression and shear.

Biomechanical Phenomena↗
PubMed2003

Advances in methodology for the validation of methods according to the International Organization for Standardization. Application to the determination of benzoic and sorbic acids in soft drinks by high-performance liquid chromatography.

Robust chemometric techniques such as least median of squares regression, H15 Huber estimator and Lenth's method are fundamental tools in the validation of analytical methods since they contribute the strategies needed to estimate efficiently parameters such as robustness, linear range, selectivity, accuracy (trueness and precision) and the capability of detection. In addition, the capability of discrimination defined as a generalisation of the capability of detection for any nominal concentration is evaluated. The new strategy proposed is applied to the validation of a chromatographic method for use in systematic analysis.

Benzoic Acid↗
PubMed1999

Application of a peptide bank from porcine brain in isolation of regulatory peptides.

Over the past years, the introduction of biological assay systems, random peptide sequencing and orphan receptor screening has led to the isolation and identification of new regulatory peptides with potential clinical impact. We have developed a method for separating peptides into about 300 fractions from large amounts of porcine brain tissue. The preparation of this peptide bank consists of three steps including ultrafiltration followed by cation-exchange separation and reversed-phase chromatography. These fractions represent the peptide bank with desalted and lyophilized peptides from brain tissue. Molecular masses of the peptides in the fractions are determined by matrix-assisted laser desorption ionization MS and a mass data bank is subsequently generated. For systematic analysis of the peptides, a subsequent two-step purification procedure is followed by Edman sequencing resulting in the identification of different peptides. A survival assay with a neuronal cell line revealing the stimulatory and inhibitory activities is applied as a model to test the 300 fractions. This primary screen indicates that the biological activities of the extracted peptides are easily characterized and, moreover, can be related to the biochemical entities. We conclude that the established peptide bank is an efficient and useful tool for the isolation of regulatory brain peptides applying different purification strategies.

Animals↗
PubMed2002

Alternate usage of two dimerization initiation sites in HIV-2 viral RNA in vitro.

An essential step in the replication cycle of all retroviruses is the dimerization of genomic RNA prior to or during encapsidation and budding. In HIV-1, a stem-loop structure in the genomic RNA called the dimerization initiation site, or DIS, has been well characterized. However, the identification of the structure(s) necessary for dimerization of HIV-2 genomic RNA has been less straightforward, as reflected by recent conflicting reports. Here, using a variety of mutant and wild-type RNA constructs and a systematic analysis of experimental conditions, we demonstrate that two dimerization sites in HIV-2 RNA are clearly discernible under different experimental conditions. A short sequence overlapping the primer binding site acts as the default dimerization site for wild-type viral RNA transcripts of several lengths provided that dimerization incubation conditions do not include a high heat step (>50 degrees C), and electrophoresis is carried out under mild conditions that do not deplete the RNA of magnesium. However, some RNA constructs are able to dimerize through stem-loop 1 (SL1), which is the structure homologous to the HIV-1 DIS, under certain experimental conditions. Interestingly, deletion or mutation of the default PBS dimerization site leads to efficient usage of the SL1 dimerization site. This study defines conditions under which each site may be used for dimerization and demonstrates, furthermore, the facility with which the two sites can substitute for each other. This is suggestive of a switching mechanism that may be used in the viral replication cycle.

Base Sequence↗
PubMed1998

Tracheal occlusion in the fetal rat: a new experimental model for the study of accelerated lung growth.

BACKGROUND: Prenatal tracheal occlusion accelerates fetal lung growth, but the mechanism of this phenomenon is unknown. Previous animal models have been limited by expense, lack of species-specific molecular probes, or the stage of lung development when studies could be performed. To provide a model that is more amenable to systematic analysis, we have developed an in vivo rat model of prenatal tracheal occlusion. METHODS: Time-dated pregnant rats underwent laparotomy at 19 days' gestational age (term, 22 days). The fetal head and neck were exteriorized through a hysterotomy, and the trachea was ligated under a dissecting microscope. The fetus was returned to the amniotic cavity, and the uterine and maternal abdominal incisions were closed. The dam and the fetuses were killed at 21.5 days' gestational age, and the fetal lungs were assessed for lung growth and compared with nonoperated littermate controls. RESULTS: Thirty-two of 50 manipulated fetuses survived. Of the 32 survivors, successful tracheal ligation was confirmed in 20, and these 20 fetuses were compared with 33 littermate controls. Fetal body weight (4.81+/-0.26 g v 4.87+/-0.41 g) and heart weight (0.05+/-0.01 g v 0.05+/-0.01 g) were not significantly different between ligated fetuses and littermate controls, whereas the wet lung weight (0.30+/-0.06 g v 0.13+/-0.02 g, P<.01), lung-to-body-weight ratio (6.34+/-1.16% v 2.64+/-0.41%, P<.01), dry lung weight (17.4+/-2.45 mg v 12.1+/-1.87 mg, P<.01), total lung DNA (1210+/-371 microg v 828+/-208 microg, P<.01), and total lung protein (14.3+/-5.3 mg v 8.7+/-1.7 mg, P<.01) were increased significantly in the ligated fetuses. The enlarged lung demonstrated normal histology findings after inflation fixation. CONCLUSIONS: Prenatal tracheal occlusion during the canalicular stage of lung development accelerates lung growth in the rat. In comparison with other large animal models, this relatively inexpensive small animal model has the distinct advantages of a short gestation, a large number of fetuses per litter, the availability of a developmental model of congenital diaphragmatic hernia, and the availability of well-defined molecular probes to investigate the mechanism of tracheal occlusion-induced lung growth.

Animals↗
PubMed1993

Staging systems of lung cancer.

The staging of lung cancer involves assessment of the anatomic extent of disease based on the best available data. Such a definition of neoplastic burden facilitates the systematic analysis and meaningful communication of diagnostic, therapeutic, and prognostic information. Clinical staging involves the best estimate of extent of disease before performance of surgical resection or biopsy procedures (or both). Surgical-pathologic staging is based on the histopathologic analysis of resected specimens, including determining the extent of local and regional disease. During the past 50 years, two major classification schemes for staging of lung cancer have evolved--one for non-small-cell lung cancers (the TNM system, indicating the status of primary tumor [T], regional lymph node [N], and metastatic [M] involvement) and the other for small-cell carcinoma of the lung (based on limited versus extensive disease). In this report, we review the evolution of the current staging systems used for primary lung cancer and their prognostic implications.

Carcinoma, Non-Small-Cell Lung↗
PubMed2004

[Bed rest or early mobilization as treatment of deep vein thrombosis: a systematic review and meta-analysis].

BACKGROUND AND OBJECTIVE: We aimed to know if treatment of deep vein thrombosis (DVT) with early mobilisation is as safe and effective as bed rest. MATERIAL AND METHOD: MEDLINE, EMBASE, Cochrane library (CCTR), Spanish Medical Index, and MD-Consult Virtual Library databases were searched. We also cross-checked bibliographies of the retrieved articles. The TESEO database of doctoral theses in Spain was also revised. We only searched for clinical trial articles comparing bed rest with early mobilization with respect to the incidence of objectively diagnosed pulmonary embolism (PE) in patients treated for DVT of lower limbs. The concordance coefficient was evaluated by statistical methods. We used relative risk and 95% confidence intervals. Selection bias was evaluated using funnel plot. RESULTS: Only three articles were included in the metaanalysis, with 296 patients randomized from 773 patients initially evaluated, with a follow-up of 9 days to 3 months. Quality rating ranged from 61.4 to 90% and the kappa index of concordance ranged from 0.78 to 0.93. The relative risks of PE between the two groups of treatment (early mobilization versus bed rest) were 1.31 (0.63-2.72), 1.50 (0.17-13.23), and 1.45 (0.56-3.75), respectively, and the global RR was 1.37 (0.78-2.40). CONCLUSIONS: The analyzed studies reveal that the treatment of DVT with early mobilization rather than bed rest neither increases the rate of PE nor increases the complication rate. New well designed, controlled clinical trials are needed to confirm the conclusions of this review.

Bed Rest↗
PubMed1998

Large scale ENU screens in the mouse: genetics meets genomics.

The worldwide effort to completely sequence the human and mouse genome will be accomplished within the next years. The focus of current activities within the framework of human genome research is mainly on the assembly of high resolution genetic and physical maps and genomic sequencing. Cloning of new genes is getting more easy using those maps. Nevertheless, it is necessary to work on a systematic analysis of gene function. Results obtained from these efforts will be of enormous value for future biological and biomedical research. However, even the complete sequence will not in all cases reveal the molecular and cellular role of the different genes. Therefore, the next phase of the Human Genome Project will have at its core the functional analysis of genes. Those genes relevant for the diagnosis, prevention and therapy of human diseases are of particular interest. Looking at the history of life sciences, mutants have been the most important tool to obtain insight into the biological function of genes. The mouse is the model of choice for the study of inherited diseases in man. In order to meet the requirements for functional human genome analysis, we need a large number of mouse mutants similar to the collection of mutants available for other model organisms such as flys and worms. To fully apply the power of genetics, multiple alleles of the same gene such as hypomorphs or hypermorphs are required. Efficient production of mouse mutants showing specific phenotypes can be achieved by the use of ethylnitrosourea (ENU). ENU is the most powerful mutagen known and we currently see a renaissance of ENU mutagenesis. The application of ENU mutagenesis is reviewed and discussed in the context of a new era of functional genomics.

Animals↗
PubMed1997

Contrasting biophysical and pharmacological properties of T-type and R-type calcium channels.

In contrast to other kinds of voltage-gated Ca2+ channels, the underlying molecular basis of T-type and R-type channels is not well-understood. To facilitate comparisons with cloned Ca2+ channel subunits, we have carried out a systematic analysis of the properties of T-type currents in undifferentiated NG108-15 cells and R-type currents in cerebellar granule neurons. Marked differences were found in their biophysical and pharmacological features under identical recording conditions. T-type channels became activated at potentials approximately 25 mV more negative than R-type channels; however, T-type channels required potentials approximately 15 mV less negative than R-type channels to be available. Accordingly, T-type channels display a much larger overlap between the curves describing inactivation and activation, making them more suitable for generating sustained Ca2+ entry in support of secretion or pacemaker activity. In contrast, R-type channels are not equipped to provide a steady current, but are very capable of supplying transient surges of Ca2+ influx. In response to a series of increasingly strong depolarizations T-type and R-type Ca2+ channels gave rise to very different kinetic patterns. T-type current records crossed each other in a characteristic pattern not found for R-type currents. These biophysical distinctions were independent of absolute membrane potential and were, therefore, complementary to the conventional categorization of T- and R-type Ca2+ channels as low- and high-voltage activated. R-type channels deactivated approximately eight-fold more quickly than T-type channels, with clear consequences for the generation of divalent cation influx during simulated action potentials. Pharmacological comparisons revealed additional contrasts. R-type current was responsive to block by omega-Aga IIIA but not nimodipine, while the opposite was true for T-type current. Both channel types were potently inhibited by the non-dihydropyridine compound mibefradil. In all respects examined, R-type currents were similar to currents derived from expression of the alpha1E subunit whereas T-type currents were not.

Animals↗
PubMed2003

Labor induction versus expectant management for postterm pregnancies: a systematic review with meta-analysis.

OBJECTIVE: To compare routine labor induction with expectant management for patients who reach or exceed 41 weeks' gestation. DATA SOURCES: Computerized databases, references in published studies, and textbook chapters in all languages were used to identify randomized controlled trials (RCTs) evaluating induction and expectant management of labor for postterm pregnancies. METHODS OF STUDY SELECTION: We identified RCTs that compared induction and expectant management for uncomplicated, singleton, live pregnancies of at least 41 weeks' gestation and evaluated at least one of the following: perinatal mortality, mode of delivery, meconium-stained fluid, meconium aspiration syndrome, meconium below the cords, fetal heart rate (FHR) abnormalities during labor, cesarean deliveries for FHR abnormalities, abnormal Apgar scores, and neonatal intensive care unit (NICU) admissions. The primary outcomes assessed were cesarean delivery rate and perinatal mortality. TABULATION, INTEGRATION, AND RESULTS: Sixteen studies met inclusion criteria for this review. For each study with binary outcomes, an odds ratio (OR) with 95% confidence intervals (CIs) was calculated for selected outcomes. Estimates of ORs for dichotomous outcomes were calculated using fixed and random-effects models. Homogeneity was tested across the studies. Compared with women allocated to expectant management, those who underwent labor induction had lower cesarean delivery rates (20.1% versus 22.0%) (OR 0.88; 95% CI 0.78, 0.99). Although subjects whose labor was induced experienced a lower perinatal mortality rate (0.09% versus 0.33%) (OR 0.41; 95% CI 0.14, 1.18), this difference was not statistically significant. Similarly, no significant differences were noted for NICU admission rates, meconium aspiration, meconium below the cords, or abnormal Apgar scores. CONCLUSION: A policy of labor induction at 41 weeks' gestation for otherwise uncomplicated singleton pregnancies reduces cesarean delivery rates without compromising perinatal outcomes.

Cesarean Section↗
PubMed2003

Prenatal bladder drainage in the management of fetal lower urinary tract obstruction: a systematic review and meta-analysis.

OBJECTIVE: To estimate the effect of prenatal bladder drainage on perinatal survival in fetuses with lower urinary tract obstruction. DATA SOURCES: Relevant articles were identified by searching the databases MEDLINE (1966-2002), EMBASE (1988-2002), and the Cochrane library (2000;4). METHODS OF STUDY SELECTION: Studies were selected if the effect of prenatal bladder drainage (vesicocentesis, vesicoamniotic shunt, or open fetal bladder surgery) on perinatal survival was reported in fetuses with ultrasonic evidence of lower urinary tract obstruction. Study selection, quality assessment, and data abstraction were performed independently and in duplicate. TABULATION, INTEGRATION, AND RESULTS: Sixteen observational studies that included nine case series (147 fetuses) and seven controlled series (195 fetuses) were identified. Study characteristics and quality were recorded for each study. Data on the effect of bladder drainage on perinatal survival were abstracted. Where controlled data were available, 2 x 2 tables were generated to compare the effects of bladder drainage versus no bladder drainage on perinatal survival. Pooled odds ratios (ORs) were used as summary measures of effect, and the results were stratified according to predicted fetal prognoses (based on ultrasound features and fetal urinary electrolytes). Among controlled studies, bladder drainage appeared to improve perinatal survival relative to no drainage (OR 2.5; 95% confidence interval [CI] 1.1, 5.9; P =.03). However, this observation was largely because among the subgroup of fetuses with a poor prognosis there was a marked improvement (OR 8.1; 95% CI 1.2, 52.9; P =.03). Improved perinatal outcome was also suggested in those fetuses considered to have a good prognosis (OR 2.8; 95% CI 0.7, 10.8; P =.13). CONCLUSION: There is a lack of high quality evidence to reliably inform clinical practice regarding prenatal bladder drainage in fetuses with ultrasonic evidence of lower urinary tract obstruction. The limited available evidence suggests that prenatal bladder drainage may improve perinatal survival in these fetuses, particularly those with poor predicted prognoses. Further research in the form of a multicenter randomized controlled trial is required to assess the short- and long-term effects of this intervention.

Drainage↗
PubMed2004

[Subjective evaluation of surgical treatment for patellar instability].

PURPOSE OF THE STUDY: We analyzed functional outcome after surgical treatment for patellar instability. MATERIAL AND METHODS: This study included 130 patients (174 knees) treated between 1988 and 1999. Minimum follow-up after surgery was two Years, mean five years (range 24-152 months). The IKDC 1999 subjective evaluation chart was used. This chart has ten items to establish level of sports activity and functional status of the knee for daily life activities. One hundred ten patients (84.5%) responded to the questionnaire. RESULTS: Eighty-eight patients were seen for clinical assessment and 22 were questioned by telephone: 104 patients (94.5%) were satisfied or very satisfied, five (4.5%) were dissatisfied, and one was disappointed. We evaluated results on the basis of pain (37.6% with climate-related pain or discomfort), residual effusion, and sensation of blocked knee (15.8%), as well as daily activities (68% with discomfort when kneeling), and sports activities (with level of activity). DISCUSSION: The patients' subjective assessment showed that surgical treatment with medialization and/or lowering of the anterior tibial tuberosity is effective with a good or excellent rate of satisfaction. The quality of these results was directly related to careful individualization of the lesions with systematic analysis of factors leading to patellar instability (trochlear dysplasia, patellar height, quadriceps dysplasia, TA-GT measures, length of the patellar tendon).

Activities of Daily Living↗
PubMed2004

The environmental occurrence of hexabromocyclododecane in Sweden.

The brominated flame retardant hexabromocyclododecane (HBCD) is extensively used in Europe, but data on the environmental concentrations of this chemical are scarce. A first screening has been performed concerning the environmental occurrence of HBCD in Sweden, a country where the chemical is not produced and the current industrial use is very limited. Possible emission sources were identified through a systematic analysis of the use in a life cycle perspective. In addition to a few point sources, diffuse emissions from polymeric products are possible. Measurements have been performed close to certain possible point sources, in the urban environment and in remote regions, and included air, deposition, water, soil, sediments, sludge, biota and foodstuffs. HBCD was detected in all media analysed and in all environments. The relatively high concentrations detected in herring and foodstuffs provide evidence for bioaccumulation of HBCD. The presence of HBCD in remote background air implies that HBCD has potential for long-range atmospheric transport. There are also some indications that diffuse emissions of HBCD occur in the urban environment.

Animals↗
PubMed1998

Effect of age on mitogen induced protein tyrosine phosphorylation in human T cell and its subsets: down-regulation of tyrosine phosphorylation of ZAP-70.

Several events of T cell activation have been reported to decline in humans with age. Since protein tyrosine phosphorylation is an early critical event of T cell activation, we performed a systematic analysis of the age-associated changes in the mitogen induced protein tyrosine phosphorylation of human T lymphocytes using SDS-PAGE and Western blotting techniques. Following stimulation with Con A and PHA, an identical pattern of protein tyrosine phosphorylation was observed in the lysates of T cells prepared from seven healthy young adults and eight healthy elderly human subjects. Five different high molecular mass proteins (75, 115, 120, 140 and 170 kDa) were consistently tyrosine phosphorylated in all of the donors from both age groups and peaked between 3 and 10 min. Tyrosine phosphorylation of the above substrates was observed in both CD4 and CD8 subsets. When compared for individual donors from both age groups, variations in the T cell response with regard to net tyrosine phosphorylation for all the substrates was observed. However, the mitogen induced level of tyrosine phosphorylation of only p75 was found to be significantly lower in unfractionated T cells as well as CD4 and CD8 subsets of older subjects than that of young subjects. Using immunoblotting, p75 was identified as ZAP-70, a member of the syk family of protein tyrosine kinases. Understanding of the biochemical basis of the reduced level of tyrosine phosphorylation of ZAP-70 will be helpful in delineating the molecular basis of age-associated impairment of T cell activation.

Adult↗
PubMed2006

Beta-rolls, beta-helices, and other beta-solenoid proteins.

Beta-rolls and beta-helices belong to a larger group of topologically similar proteins with solenoid folds: because their regular secondary structure elements are exclusively beta-strands, they are referred to as beta-solenoids. The number of beta-solenoids whose structures are known is now large enough to support a systematic analysis. Here we survey the distinguishing structural features of beta-solenoids, also documenting their notable diversity. Appraisal of these structures suggests a classification based on handedness, twist, oligomerization state, and coil shape. In addition, beta-solenoids are distinguished by the number of chains that wind around a common axis: the majority are single-stranded but there is a recently discovered subset of triple-stranded beta-solenoids. This survey has revealed some relationships of the amino acid sequences of beta-solenoids with their structures and functions-in particular, the repetitive character of the coil sequences and conformations that recur in tracts of tandem repeats. We have proposed the term beta-arc for the distinctive turns found in beta-solenoids and beta-arch for the corresponding strand-turn-strand motifs. The evolutionary mechanisms underlying these proteins are also discussed. This analysis has direct implications for sequence-based detection, structural prediction, and de novo design of other beta-solenoid proteins. The abundance of virulence factors, toxins and allergens among beta-solenoids, as well as commonalities of beta-solenoids with amyloid fibrils, imply that this class of folds may have a broader role in human diseases than was previously recognized. Thus, identification of genes with putative beta-solenoid domains promises to be a fertile direction in the search for viable targets in the development of new antibiotics and vaccines.

Models, Molecular↗
PubMed1995

Studies on protein stability with T4 lysozyme.

A series of mutants of phage T4 lysozyme have been constructed and have permitted a systematic analysis of different aspects of protein stability and folding. One striking result has been the ability of the protein to accommodate changes at many sites, yet still fold and retain activity. This shows that many amino acids in the sequence of the protein are nonessential for protein folding and stability. Such amino acids appear, predominantly, to be those that are mobile and/or largely exposed to solvent. Even though buried residues seem to be more important, it is still possible to change these as well. In some cases individual amino acids have been replaced. In other cases it has been found that a combination of substitutions permits repacking of the core. Such repacking is associated with adjustments of both the main chain and side chains. Only rarely do side chains rotate into radically new orientations. Taken together, substitutions of core residues confirm the overall importance of the hydrophobic effect as the dominant factor in stabilizing the folded structures of proteins. "Cavity-creating" substitutions of the form Leu-->Ala show, as expected, that the burial of the bulky leucine side chain within the core of the protein confers greater hydrophobic stabilization than is the case for the smaller alanine side chain. Leu-->Ala substitutions that create large cavities are especially destabilizing because they result in a loss of both hydrophobic and van der Waals interactions. In cases where the protein relaxes to reduce the size of the putative cavity, alternative van der Waals interactions are generated and the overall destabilization of the protein may be less severe than in cases where a large cavity is formed.

Amino Acid Sequence↗
PubMed1998

Rhodopsin: a prototypical G protein-coupled receptor.

A variety of spectroscopic and biochemical studies of recombinant site-directed mutants of rhodopsin and related visual pigments have been reported over the past 9 years. These studies have elucidated key structural elements common to visual pigments. In addition, systematic analysis of the chromophore-binding pocket in rhodopsin and cone pigments has led to an improved understanding of the mechanism of the opsin shift, and of particular molecular determinants underlying color vision in humans. Identification of the conformational changes that occur on rhodopsin photoactivation has been of particular recent concern. Assignments of light-dependent molecular alterations to specific regions of the chromophore have also been attempted by studying native opsins regenerated with synthetic retinal analogs. Site-directed mutagenesis of rhodopsin has also provided useful information about the retinal-binding pocket and the molecular mechanism of rhodopsin photoactivation. Individual molecular groups have been identified to undergo structural alterations or environmental changes during photoactivation. Analysis of particular mutant pigments in which specific groups are locked into their respective "off" or "on" states has provided a framework to identify determinants of the active conformation, as well as the minimal number of intramolecular transitions required to switch between inactive and active conformations. A simple model for the active state of rhodopsin can be compared to structural models of its ground state to localize chromophore-protein interactions that may be important in the photoactivation mechanism. This review focuses on the recent functional characterization of site-directed mutants of bovine rhodopsin and some cone pigments. In addition, an attempt is made to reconcile previous key findings and existing structural models with information gained from the analysis of site-directed mutant pigments.

Amino Acid Sequence↗