solubility and stability engineering of human vh domains

... activity of the human enzyme was not regained upon removal of EDTA and addition of metal-ions Figure shows the hydrophobicity profiles of human and P fluorescens ACMSD, performed according to Kyte and ... [21] From the sequence alignment of human and bacterial ACMSD and the comparison of their three-dimensional structures, the presence of the metallocofactor in the human enzyme can be reliably predicted ... the mean of three independent experiments and are presented as copies of the target variant per 109 copies of 18S RNA Quantitation of ACMSD variants in liver, kidney and brain An isoform-specific...

Ngày tải lên: 19/02/2014, 02:20

... [20]: koff ẳ koff2ị ẵibuprofen=K2 ẵibuprofenị ỵ koff3ị ẵibuprofen=K3 ẵibuprofenịị ỵ kỵ off 7ị where koff(2) and koff(3) indicate values of koff occurring at K2 < [ibuprofen] < K3, and at ... functional and structural properties of domains I and II [i.e of the 1382 region of (t)HSA] l The values of the Hill coefcients (n1, n2, and n3) for ibuprofen binding to tHSAheme and HSAheme ... low-afnity sites of tHSAheme-Fe(III) and HSAheme-Fe(III) [represented by a and (1 a), respectively, in Eqn (1)] are 0.18 and 0.82, and 0.14 and 0.86, respectively (Table 1) Effect of ibuprofen on peroxynitrite...

Ngày tải lên: 06/03/2014, 01:20

... the development of the advanced functions of human nervous system in light of the Gene Ontology (GO) profile of human orthologs Results Characters and distribution of LIRs in human autosomes We ... rhesus monkey and mouse orthologs Our data suggest that most of the LIRs shared by human and chimpanzee orthologs were developed and maintained by the common ancestor of humans and chimpanzees ... large number of (CA)n and (GA)n repeats in the human genome, the frequency of their complementary repeats (TG)n and (CT)n is much lower [37] Therefore, the presence of the TG ⁄ CA-rich and TC ⁄ GA-rich...

Ngày tải lên: 07/03/2014, 00:20

... properties of SEPT2 for GTPcS and GDP, respectively, may also indicate different conformations in recognizing triphosphate and diphosphate guanine nucleotides The similarity of the Kd, koff and kcat of ... Huang et al GTPase properties of human SEPT2 Table Summary of the GTPcS and GDP dissociation constants and stoichiometrical binding site (n) for wild-type SEPT2 and S218A mutant The data for ... [17] and yeast [22] Complexes comprised of different septin isoforms would reflect the binding and hydrolysis properties of each protein Indeed, it has been reported that yeast septins Cdc10 and...

Ngày tải lên: 07/03/2014, 12:20

... chromatography and chromatofocusing, as described previously [21] Results Expression and secretion of truncated variants of GCPII To analyze the contribution of individual domains of human GCPII ... NaCl and 20 lL of the GCPII sample, were preincubated for 15 at 37 °C in a final volume of 225 lL A 25 lL mixture of 950 nM ÔcoldÕ NAAG (Sigma) and 50 nM H-labelled NAAG (51.9 CiÆmmol)1; New England ... domain (domain E) and the D domain of rat GCPII The contribution of domains C and F to the GCPII hydrolytic activity/inhibitor binding remains to be established The 3D structure of GCPII has not...

Ngày tải lên: 07/03/2014, 15:20

... Amino-acid sequence and tryptic peptides of human HCII The peptides indicated below were detected by MALDI-TOF- and/ or ESI-MS (see Figs and 2, respectively) in tryptic digests of human HCII isolated ... were isolated at random, exposed to 0.05 lM MTX, expanded, and checked for the presence of human HCII mRNA by RT-PCR Positive cell lines were picked and after several rounds of selection with ... properties of HCII Here, we report on the structure of posttranslational modifications of plasma-derived and recombinant human HCII from CHO cells and their effects on heparin binding MATERIALS AND METHODS...

Ngày tải lên: 08/03/2014, 22:20

... composed mainly of a helices (Figs and 2) The core of Iba2 is a pair of EF-hand motifs, denoted as EF-hands and 2, each consisting of two a helices (aA, aB and aC, aD, respectively) anking a ... cross-linking efciency of Iba1 and Iba2 or in the overall morphology of the generated lament bundles Calcium afnity of Iba1 and Iba2 Homodimerization and actin binding of Iba1 and Iba2 were similar ... similarities and differences between Iba1 and Iba2 We investigated Ca2+ binding and homodimerization of Iba1 and Iba2 Furthermore, F-actin binding and cross-linking assays were performed with both human...

Ngày tải lên: 16/03/2014, 06:20

... 400 nm with excitation and emission band pass of nm and nm, respectively ð3Þ DGUÀFH2 O ð4Þ where aF and aU are the intercepts and bF and bU are the slopes of the baselines of the equilibrium fluorescence ... of Nile red Ó FEBS 2004 Mechanism of folding and unfolding of HSA (Eur J Biochem 271) 1795 Fig Equilibrium refolding of HSA in the presence of different concentrations of GdnHCl The recovery of ... values of m and Dm The curve fittings were performed using MICROSOFT ORIGIN software Calculation of the rate constants from the kinetics of refolding HSA was found to follow two phases of refolding,...

Ngày tải lên: 16/03/2014, 16:20

... and the Role of Colonic Bacteria 76 89 Digestion a n d A b s o r p t i o n of Carbohydrate 91 Gary M Gray, M.D Carbohydrate Components of the Human Diet 92 Sites and Mechanisms of Digestion of ... Digestion, and Absorption Transport and Conversion of Riboflavin to Coenzymes 469 471 xxii • • • Contents Riboflavin Catabolism and Excretion 472 Functions of Flavocoenzymes in Metabolism 472 Riboflavin ... Assessment of Riboflavin Nutriture 475 Riboflavin Requirements 475 Thiamin Thiamin and Thiamin Coenzyme Structure and Nomenclature Sources, Digestion, and Absorption 476 Transport and Conversion of Thiamin...

Ngày tải lên: 16/03/2014, 18:07

... Time course of excision of 8-oxoGua (d), FapyGua (s) and FapyAde ( ) by OGG1 (B) Time course of excision of 8-oxoGua (d), FapyGua (s) and FapyAde ( ) by Fpg Means and standard deviations of three ... values of kinetic constants for excision of FapyGua and 8-oxoGua by various forms of OGG1 are 500 400 300 200 100 0 10 15 20 Time (min) Fig Excision of 8-oxoGua and FapyGua by wild-type OGG1 and ... values of kcat and Km for FapyGua excision were Table Km, kcat and ksp values for excision of FapyGua and 8-oxoGua from c-irradiated calf thymus DNA by wild-type and mutant OGG1 proteins Mean ± standard...

Ngày tải lên: 23/03/2014, 05:22

Ngày tải lên: 23/03/2014, 05:23

... is human Hsp90a (PP30[hHsp90a]; see Experimental procedures) Western blotting using an antiserum that recognizes, with comparable efficiencies, both of the yeast and both of the human isoforms of ... phosphorylation of the MAP kinase activation loop [3,14,18], Y2H interaction of these MAP kinases with the two isoforms of human Hsp90 was strengthened by heat shock (Fig 5A) The stronger interaction of ERK5 ... Hsp90, with altered cochaperone function and with the loss of plasma membrane drug efflux pumps [35] The results of this study point to the two isoforms of human cytosolic Hsp90 differing in the...

Ngày tải lên: 23/03/2014, 07:20

... islands with a CGobs/CGexp ratio in excess of 0.6 and a G + C content of 62% spanning two regions from )377 to )158 and from )137 to +136 of the start codon Such a CpG island is indicative of ... and PLP has an extinction coefficient of 5900 M)1Æcm)1 One unit of specific activity is defined as the amount of protein that catalyses the formation of lmol PLPÆmin)1 at 25 °C The value of Km and ... (left) and the 3¢-UTR of human PNPO cDNA (right) The membrane was stripped and reprobed with a b-actin cDNA probe (bottom) The approximate sizes of the isoforms are indicated (B) The scheme of two...

Ngày tải lên: 23/03/2014, 12:20

... splicing of PP17 variants, the importance of protein kinase A (PKA)- and protein kinase C (PKC)dependent pathways for the regulation of PP17 gene expression was demonstrated, and the effect of the ... structure and regulation A GenBank search revealed a high variety of alternatively spliced human ESTs ) related to PP17a and PP17b cDNAs by length and sequence ) in almost all types of healthy ... kb, 15q26) and intron sizes were divergent, homology was proven by the similar number and length of exons, the corresponding analogous peptide lengths, and the high number of identical and conserved...

Ngày tải lên: 23/03/2014, 21:20

... public and private domains fuel questions about the interpretation and applicability of current policies governing the conduct of social and behavioral research involving human subjects The Office ... course of one -and- a-half days, they fleshed out the relevant issues in online research and considered the role of IRBs This report and its action, research and education agenda are products of the ... others This lack of understanding by participants, and sometimes researchers as well, of the technical and storage capabilities of Internet technologies may elevate the risk The risk of exposure can...

Ngày tải lên: 29/03/2014, 20:20

... of open and closed models of human P-gp Table Amino acids involved in the binding cavity of the inward (‘in’) and outward (‘out’)-facing conformations of human P-gp; the numbers and letters of ... Comparison of open and closed models of human P-gp A C B D Fig The interaction panel of the ligand QZ59-SSS in the X-ray structure of mouse P-gp (PDB ID code 3G61): (A) ligand 1, molecule A; (B) ligand ... Tyr303 (human Tyr307) Ligand (molecule A) and ligand (molecule B) were not fully resolved and the analyses of the ligand interactions thus involved only parts of the structures For ligand (Fig...

Ngày tải lên: 29/03/2014, 22:21

... be attributed to an anisotropy of the porphyrin absorption band [22] Upon addition of substrate, the signal of the positive CD band at 290 nm and the negative band at 386 nm decreased (Fig 4B) ... standard After extraction of the steroids and evaporation of the chloroform phase, the steroids were resuspended in 200 lL acetonitrile and separated on a Jasco reversed phase HPLC system of ... JM109 system used for the expression of rat CYP11B1 and CYP11B2 [15] The expressed form of hCYP11B1 was stable in the presence of detergents and glycerol and highly purified through three chromatographic...

Ngày tải lên: 30/03/2014, 04:20

... effect of exercise Now let’s suppose Sam goes on the diet D and the exercise regime E If Sam gets the benefits of both—the main effect of D and the main effect of E—then the interaction of D and ... proper models outperform humans This picture of humans ‘‘calculating’’ suboptimal formulas, of implicitly using improper models, 30 Epistemology and the Psychology of Human Judgment also fits with ... the success of proper linear models for problems of human and social prediction gets for free the explanation of the success of improper linear models That’s because for certain kinds of problem,...

Ngày tải lên: 10/06/2014, 22:00

... kind permission of Professor Warwick and of icube.co.uk CYBORGS UNPLUGGED 19 Fig 1.4 Diagram of implant used by Professor Kevin Warwick By kind permission of Professor Warwick Each of the 100 tips ... sense of combining flesh and wires but in the more profound sense of being human- technology symbionts: thinking and reasoning systems whose minds and selves are spread across biological brain and ... of deep human- machine symbiosis really expand and alter the shape of the psychological processes that make us who we are The old technologies of pen and paper have deeply impacted the shape and...

Ngày tải lên: 11/06/2014, 00:24

... than that of other primates in terms of the range of sound types we can produce Many basic varieties of vowels and consonants are beyond the articulatory capacity of nonhumans, or of earlier ... understanding of the syntax of human natural languages Research focused almost exclusively on the systematic role of sound in language and on the structure of words Only with the development of generative ... Consider the definition of humans as ‘‘featherless bipeds.’’ Who would be content with that as an expression of the nature of humanness (or of primateness, if you think of chimpanzees and gorillas as...

Ngày tải lên: 11/06/2014, 14:43