Common structural features of cholesterol binding sites in crystallized soluble proteins

Cholesterol-protein interactions are essential for the architectural organization of cell membranes and for lipid metabolism. While cholesterol-sensing motifs in transmembrane proteins have been identified, little is known about cholesterol recognition by soluble proteins. We reviewed the structural characteristics of binding sites for cholesterol and cholesterol sulfate from crystallographic structures available in the Protein Data Bank. This analysis unveiled key features of cholesterol-binding sites that are present in either all or the majority of sites: i) the cholesterol molecule is generally positioned between protein domains that have an organized secondary structure; ii) the cholesterol hydroxyl/sulfo group is often partnered by Asn, Gln, and/or Tyr, while the hydrophobic part of cholesterol interacts with Leu, Ile, Val, and/or Phe; iii) cholesterol hydrogen-bonding partners are often found on α-helices, while amino acids that interact with cholesterol's hydrophobic core have a slight preference for β-strands and secondary structure-lacking protein areas; iv) the steroid's C21 and C26 constitute the “hot spots” most often seen for steroid-protein hydrophobic interactions; v) common “cold spots” are C8–C10, C13, and C17, at which contacts with the proteins were not detected. Several common features we identified for soluble protein-steroid interaction appear evolutionarily conserved.     

A framework for exhaustively mapping functional missense variants

Although we now routinely sequence human genomes, we can confidently identify only a fraction of the sequence variants that have a functional impact. Here, we developed a deep mutational scanning framework that produces exhaustive maps for human missense variants by combining random codon mutagenesis and multiplexed functional variation assays with computational imputation and refinement. We applied this framework to four proteins corresponding to six human genes: UBE2I (encoding SUMO E2 conjugase), SUMO1 (small ubiquitin‐like modifier), TPK1 (thiamin pyrophosphokinase), and CALM1/2/3 (three genes encoding the protein calmodulin). The resulting maps recapitulate known protein features and confidently identify pathogenic variation. Assays potentially amenable to deep mutational scanning are already available for 57% of human disease genes, suggesting that DMS could ultimately map functional variation for all human disease genes.     

Exploring the expression bar code of SAA variants for gastric cancer detection

We reported an integrated platform to explore serum protein variant pattern in cancer and its utility as a new class of biomarker panel for diagnosis. On the model study of serum amyloid A (SAA), we employed nanoprobe‐based affinity mass spectrometry for enrichment, identification and quantitation of SAA variants from serum of 105 gastric cancer patients in comparison with 54 gastritis patients, 54 controls, and 120 patients from other cancer. The result revealed surprisingly heterogeneous and most comprehensive SAA bar code to date, which comprises 24 SAA variants including SAA1‐ and SAA2‐encoded products, polymorphic isoforms, N‐terminal–truncated forms, and three novel SAA oxidized isotypes, in which the variant‐specific peptide sequence were also confirmed by LC‐MS/MS. A diagnostic model was developed for dimension reduction and computational classification of the 24 SAA‐variant bar code, providing good discrimination (AUC = 0.85 ± 3.2E?3) for differentiating gastric cancer group from gastritis and normal groups (sensitivity, 0.76; specificity, 0.81) and was validated with external validation cohort (sensitivity, 0.71; specificity, 0.74). Our platform not only shed light on the occurrence and modification extent of under‐represented serum protein variants in cancer, but also suggested a new concept of diagnostic platform by serum protein variant profile.     

陕北榆林王阳畔遗址的植硅体分析

榆林地处农牧交错带,是研究环境演变和人地关系的理想地区。本文基于植硅体分析,探讨了该地区王阳畔遗址仰韶晚期至龙山早期过渡阶段的农业生产情况及农业对环境的适应等。遗址剖面及灰坑样品的植硅体分析表明:仰韶晚期,粟和黍是先民的主要粮食作物,而黍较粟始终保持着种植优势;龙山早期,粟作农业逐渐衰落。植硅体组合图谱显示,先民在此遗址活动期间,气候是略偏凉偏干的,整体上较为稳定,呈现出干凉化的趋势。综合来看,农业活动的强弱在一定程度上响应了环境的变化。需要指出的是,该遗址榆科植硅体的大量发现,不仅补充了史前植物利用史,同时也对丰富植硅体数据库具有重要意义。     

泥河湾盆地黑土沟遗址

黑土沟遗址是泥河湾盆地目前发现的时代较古老的一处早更新世旧石器时代考古遗址。根据磁性地层学资料判断,遗址位于Matsuyama反极性时的Olduvai正极性亚时阶段,其年龄为1.77-1.95Ma。2006年,在黑土沟遗址的考古地质勘探中,查明探坑文化层厚1.33m,由4个自然层组成;在大约7.6m~3的堆积中,出土遗物20585件,包括石制品20489件,哺乳动物骨牙碎片96件。石制品中,石核、石片、断块和器物分别占0.36%、97.90%、1.00%和0.74%,在石片中竟有87.74%的数量是碎屑。器物中出现旧石器晚期常见的圆盘状刮削器。石制品保存新鲜,发现拼合标本3组。石制品绝大部分属于微型和小型标本。砸击制品在地层中的密度较大,而且含有似棱柱状石核和似石叶薄长石片。     

Regeneration and inhibition of proton pumping activity of bacteriorhodopsin blue membrane by cationic amine anesthetics

Bacteriorhodopsin (bR) is the prototype of an integral membrane protein with seven membrane-spanning α-helices and serves as a model of the G-protein-coupled drug receptors. This study is aimed at reaching a greater understanding of the role of amine local anesthetic cations on the proton transport in the bR protein, and furthermore, the functional role of “the cation” in the proton pumping mechanism. The effect of the amine anesthetic cations on the proton pump in the bR blue membrane was compared with those by divalent (Ca²⁺, Mg²⁺ and Mn²⁺) and monovalent metal cations (Li⁺, Na⁺, K⁺ and Cs⁺), which are essential for the correct functioning of the proton pumping of the bR protein. The results suggest that the interacting site of the divalent cation to the bR membrane may differ from that of the monovalent metal cation. The electric current profile of the bR blue membrane in the presence of the amine anesthetic cations was biphasic, involving the generation and inhibition of the proton pumping activity in a concentration-dependent manner. The extent of the regeneration of the proton pump by the additives increased in the order of monovalent metal cation<monovalent amine anesthetic cation<divalent metal cation. We found that organic cations such as the amine anesthetics can also regenerate the proton pump in the bR protein. The inhibition of proton transport in the bR protein by the anesthetic cations was elucidated using the wild type, the E204Q and the D96N mutated bRs. The hydrophobic interaction of the amine anesthetics with the bR protein plays an important part in inhibiting the bR proton pump.     

Vertical Distribution of Bacterivorous Nematodes under Different Land Uses

The vertical distribution of dominant genera of bacterivorous nematodes to 150-cm depth in an aquic brown soil was compared after 14 years of four contrasting land uses, i.e., cropland-rice (CR), cropland-maize (CM), abandoned cropland (AC), and woodland (WL). The study was conducted at the Shenyang Experimental Station of Ecology, a Chinese Ecosystem Research Network (CERN) site in Northeast China. Data were analyzed using two-way analysis of variance with land use and depth as independent variables. More than 70% of Chiloplacus, Eucephalobus, and Monhystera spp. were present in the uppermost soil layer (0 to 5 cm) in the CR treatment. In contrast, Chiloplacus and Prismatolaimus spp. were distributed down to 100-cm depth in the AC and CM treatments, respectively. Differences in numbers of Acrobeles, Acrobeloides, Cephalobus, Chiloplacus, Eucephalobus, Monhystera, Plectus, and Prismatolaimus were found among land uses and at various depths. Soil C and N were correlated positively with numbers of Monhystera and Plectus in the CR treatment, Acrobeloides in the CM treatment, and Acrobeles and Acrobeloides in the AC treatment. Soil pH was correlated negatively with Monhystera, Plectus (CR), and Acrobeloides (CM, AC). The relationship of pH with Acrobeles depended on land use: positive in the WL treatment and negative in the AC treatment. Our results suggested that Cephalobus and Prismatolaimus in the CR treatment, and Chiloplacus and Prismatolaimus in the WL treatment, were insensitive to soil properties measured. Differences in vertical distribution should be considered when studying dominant bacterivorous nematode genera among land uses.     

Purification and biochemical characterization of ovine alpha-1-proteinase inhibitor: mechanistic adaptations and role of Phe350 and Met356

The glycoprotein alpha-1-proteinase inhibitor (alpha-1-PI) is a member of the serpin super family that causes rapid and irreversible inhibition of redundant serine protease activity. A homogenous preparation of ovine alpha-1-PI, a 60 kDa protein was obtained by serially subjecting ovine serum to 40-70% (NH(4))(2)SO(4) precipitation, Blue Sepharose, size-exclusion, and concanavalin-A chromatography. Extensive insights into the trypsin, chymotrypsin, and elastase interaction with ovine alpha-1-PI, point towards the involvement of Phe(350) besides the largely conserved Met(356) in serine protease recognition and consequent inhibition. The N-terminal of C-terminal peptides cleaved on interaction with elastase, trypsin, and chymotrypsin prove the presence of diffused sub-sites in the vicinity of Met(356) and the strategically positioned Pro anchored peptide stretch. Further, human alpha-1-PI is more thermolabile compared to ovine alpha-1-PI, higher thermolability is mainly attributed to poorer glycosylation. The enzymatic deglycosylation of human and ovine alpha-1-PI results in diminished thermostability of the inhibitors, with sharp decrease in thermal transition temperatures but retaining their inhibitory potency. Homology modeling of the deduced amino acid sequence of ovine alpha-1-PI using the human alpha-1-PI template has been used to explain the observed inhibitor-protease interactions.