Crystal structure of a complex between protein tyrosine phosphatase 1B and the insulin receptor tyrosine kinase

Protein tyrosine phosphatase 1B (PTP1B) is a highly specific negative regulator of insulin receptor signaling in vivo. The determinants of PTP1B specificity for the insulin receptor versus other receptor tyrosine kinases are largely unknown. Here, we report a crystal structure at 2.3 A resolution of the catalytic domain of PTP1B (trapping mutant) in complex with the phosphorylated tyrosine kinase domain of the insulin receptor (IRK). The crystallographic asymmetric unit contains two PTP1B-IRK complexes that interact through an IRK dimer interface. Rather than binding to a phosphotyrosine in the IRK activation loop, PTP1B binds instead to the opposite side of the kinase domain, with the phosphorylated activation loops sequestered within the IRK dimer. The crystal structure provides evidence for a noncatalytic mode of interaction between PTP1B and IRK, which could be important for the selective recruitment of PTP1B to the insulin receptor.     

Cloning, structural analysis, and mapping of the B30 and B7 multigenic families to the major histocompatibility complex (MHC) and other chromosomal regions

 We present the cloning, structural analysis, and mapping of new members belonging to two multigenic families, the B30-RING finger family and the B7.1-B7.2 family, as well as two genes derived by exon shuffling from members of these families. Eight new members were found and three of them map to the human major histocompatibilitiy complex (MHC) region. Phylogenic and physical mapping analysis allowed us to decipher the evolutionary story of these two multigenic families and to shed light on the evolution of the MHC region. We also show that a deductive analysis can be used to predict the existence of a given gene. Received: 17 February 1997 / Revised: 17 March 1997     

Performance, chewing activity, and ruminal parameters in yearling beef steers fed early-harvested sorghum silage: Effect of chop length and wheat straw addition

Two experiments were conducted to evaluate effects of silage chop length (SCL) and long wheat straw (WS) addition on performance, ruminal parameters, and chewing activity of yearling beef steers. The aim of the first experiment was to measure dry matter (DM) intake, average daily gain (ADG) and feed conversion (FC). For this, 59 Angus yearling steers were assigned to one of the four treatments in a 2 × 2 factorial arrangement. The factors were SCL (i.e., 5.8 and 9.4 mm mean particle length, MPL) and WS addition (i.e., 0 and 100 g/kg). Diets consisted of 792 g/kg of early-harvested sorghum silage (SS, Sorghum bicolor (L.) Moench.), 200 g/kg of pelleted sunflower meal, and 8 g/kg of urea and mineral–vitamin supplements. The amount of SS was reduced by 100 g/kg when WS was added to the diet. The second experiment, which aimed to evaluate ruminal parameters and chewing activity, consisted of two treatments with four rumen-cannulated steers fed with a diet comprised of finely chopped SS (5.8 mm MPL) either with or without WS. In the first experiment, SCL had no effect on DM intake, ADG or FC, while addition of WS depressed DM intake and ADG and had a negative effect on FC, without interactions between the factors. In the second experiment, WS addition reduced DM intake and volatile fatty acid concentration, but did not affect rumen pH or chewing activity. Results suggest that WS addition to these SS-based diets containing a high aNDF did not improve the ruminal environment, did not affect chewing activity and had a negative effect on steer performance, while SCL may not affect performance of beef steers.     

Structural basis for inhibition of the insulin receptor by the adaptor protein Grb14

Grb14, a member of the Grb7 adaptor protein family, possesses a pleckstrin homology (PH) domain, a C-terminal Src homology-2 (SH2) domain, and an intervening stretch of approximately 45 residues known as the BPS region, which is unique to this adaptor family. Previous studies have demonstrated that Grb14 is a tissue-specific negative regulator of insulin receptor signaling and that inhibition is mediated by the BPS region. We have determined the crystal structure of the Grb14 BPS region in complex with the tyrosine kinase domain of the insulin receptor. The structure reveals that the N-terminal portion of the BPS region binds as a pseudosubstrate inhibitor in the substrate peptide binding groove of the kinase. Together with the crystal structure of the SH2 domain, we present a model for the interaction of Grb14 with the insulin receptor, which indicates how Grb14 functions as a selective protein inhibitor of insulin signaling.     

A principal components analysis of the phytoplankton from a pond in the Paraná River Valley,Argentina

A principal-component scheme was applied to a set of 10 physical, chemical, and biological attributes, measured in 42 samples collected from Los Matadores pond (May 1974–July 1975). The first three components account for 82.1% of the total variance in the data and represent the lotic influence (58.3% of the variance), the seasonal cycle (16.4%), and the degree of maturity (7.4%).The authors are members of the Carrera del Investigador Cientifico y Tecnologico, Consejo Nacional de Investigaciones Cientificas y Tecnicas (CONICET), Argentina.     

Factors controlling the concentration of particulate carbohydrates and amino acids in the Paraná River

Particulate carbohydrates and particulate amino acids were determined in a set of samples from the Paraná River, Argentina, covering a stretch of over 1300 km. To gain an understanding of the factors which control the concentrations of these organic compounds, a multivariate statistical approach was used (R-mode varimax factor analysis). This technique separated the influence of various sources of organic matter. The floodplain of the middle reach (about 900 km long and 20–30 km wide) was the most significant source controlling the concentrations of particulate carbohydrates (PCH), providing angiosperm debris, phytoplankton and grasses. Particulate amino acids (PAA) concentrations were determined markedly both by contributions from the Paraguay basin, and the lotic and lentic environments of the floodplain. Sugars single out forested tributary basins as sources of bacteria while amino acids point toward the Paraguay River as a major source of biodegraded material; heterotrophic processes, however, appear more clearly depicted with PAA than with PCH. A downstream increase of the PCH/PAA ratio suggests a larger consumption of PAA in the lowermost reaches.     

Partial enzymatic deglycosylation preserves the structure of cleaved recombinant HIV-1 envelope glycoprotein trimers

The trimeric envelope glycoprotein complex (Env) is the focus of vaccine development programs aimed at generating protective humoral responses to human immunodeficiency virus type 1 (HIV-1). N-Linked glycans, which constitute almost half of the molecular mass of the external Env domains, produce considerable structural heterogeneity and are a major impediment to crystallization studies. Moreover, by shielding the peptide backbone, glycans can block attempts to generate neutralizing antibodies against a substantial subset of potential epitopes when Env proteins are used as immunogens. Here, we describe the partial deglycosylation of soluble, cleaved recombinant Env trimers by inhibition of the synthesis of complex N-glycans during Env production, followed by treatment with glycosidases under conditions that preserve Env trimer integrity. The partially deglycosylated trimers are stable, and neither abnormally sensitive to proteolytic digestion nor prone to aggregation. Moreover, the deglycosylated trimers retain or increase their ability to bind CD4 and antibodies that are directed to conformational epitopes, including the CD4-binding site and the V3 region. However, as expected, they do not react with glycan-dependent antibodies 2G12 and PGT123, or the C-type lectin receptor DC-SIGN. Electron microscopic analysis shows that partially deglycosylated trimers have a structure similar to fully glycosylated trimers, indicating that removal of glycans does not substantially perturb the structural integrity of the trimer. The glycan-depleted Env trimers should be useful for structural and immunogenicity studies.