乌司他丁联合前列地尔对急性胰腺炎的临床疗效及对血清TNF-α、IL-1β、HMGB1、HSP的影响

目的:探讨乌司他丁联合前列地尔治疗急性胰腺炎的临床疗效及对血清肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素(Interleukin-1 beta,IL-1β)、高迁移率组蛋白1(HMGB1)、热休克蛋白(HSP)的影响。方法:选择72例我院收治的急性胰腺炎患者并将其随机平均分为观察组和对照组。对照组按照常规方法治疗,观察组在常规治疗方法上采用乌司他丁联合前列地尔加入葡萄糖注射治疗。比较两组患者临床症状及体征恢复时间、治疗前后血清TNF-α、IL-1β、HMGB1、HSP水平的变化及血淀粉酶水平的恢复情况。结果:治疗后,观察组的总有效率为97.2%,对照组的总有效率为69.7%,观察组显著高于对照组(P0.05);观察组多个体征恢复时间均明显短于对照组(P0.05);两组血清TNF-α、IL-1β、HMGB1、HSP水平均较治疗前明显下降,且观察组以上指标均显著低于对照组(P0.05);观察组血淀粉酶水平恢复率明显高于对照组(P0.05)。结论:乌司他丁联合前列地尔治疗急性胰腺炎能有效提高其临床疗效,显著缩短临床症状及体征恢复时间、血淀粉酶恢复时间,降低患者血清TNF-α、IL-1β、HMGB1、HSP水平。     

清胰利胆颗粒对重症急性胰腺炎患者血清HMGB1, HSP70, HSP27,IL-8水平的影响

目的:研究清胰利胆颗粒对重症急性胰腺炎患者血清高迁移率族蛋白1(HMGB1)、热休克蛋白70(HSP70)、热休克蛋白27(HSP27)、白介素-8(IL-8)水平的影响。方法:选取2015年8月至2016年7月我院收治的84例重症急性胰腺炎患者,根据随机数字法分为观察组和对照组,42例每组。对照组采取常规方案完成治疗,观察组在此基础上使用清胰利胆颗粒治疗。比较两组患者临床疗效,血淀粉酶恢复至正常时间、白细胞恢复至正常时间、胃肠功能恢复至正常时间、腹痛缓解时间及血清HMGB1、HSP70、HSP72、IL-8水平。结果:治疗后,观察组临床总有效率显著高于对照组[92.86%(39/42)比71.43%(30/42)](P0.05)。观察组的血淀粉酶恢复至正常时间、白细胞恢复至正常时间、胃肠功能恢复至正常时间及腹痛缓解时间显著短于对照组(P0.05)。治疗前,两组患者HMGB1、HSP70、HSP72、IL-8水平比较无显著差异(P0.05),治疗后,两组患者HMGB1、HSP70、HSP72、IL-8水平和治疗前相比显著下降(P0.05),和对照组相比,观察组的HMGB1、HSP70、HSP72、IL-8水平较低(P0.05)。结论:清胰利胆颗粒能有效降低重症急性胰腺炎患者血清HMGB1、HSP70、HSP27、IL-8水平,且可提高临床疗效。     

乌司他丁辅助治疗急性重症胰腺炎的疗效及对occludin、CRPI、IL-6水平的影响

目的:研究乌司他丁辅助治疗急性重症胰腺炎的疗效及对咬合蛋白(Occludin)、C反应蛋白(快速)(CRPI)、白细胞介素6(IL-6)水平的影响。方法:选择2014年12月至2016年11月在我院进行治疗的118例急性重症胰腺炎患者,随机将其均分为观察组(59例)和对照组(59例),对照组给予常规治疗,观察组患者则在对照组治疗方案的基础上增加乌司他丁辅助治疗,两组患者的治疗疗程均为两周,记录并比较两组患者治疗前后的血清occludin、CRPI、IL-6水平,治疗后腹胀、腹痛、恶性呕吐及腹膜刺激征等体征消失时间及临床疗效。结果:治疗后,观察组和对照组患者的总有效率分别是88.1%、69.5%,观察组总有效率显著高于对照组(P0.05);观察组患者治疗后的血清occludin水平较对照组患者显著升高(P0.05),血清IL-6及CRPI水平显著低于对照组,且腹胀、腹痛、恶性呕吐、腹膜刺激征等体征消失时间显著短于对照组(P0.05)。结论:乌司他丁辅助治疗重症急性胰腺炎的临床疗效显著优于常规治疗,不仅能改善腹胀、腹痛、恶性呕吐及腹膜刺激征,而且可有效降低患者血清CRPI、IL-6水平和提高occludin水平。     

连续性血液净化治疗联合乌司他丁对急性胰腺炎患者TLR4、血浆炎症因子及白介素的影响

目的:探究连续性血液净化联合乌司他丁对急性胰腺炎患者TLR4、白介素及血浆炎症因子的影响。方法:选择2014年1月至2016年1月我院接诊的72例急性胰腺炎患者,并采用随机的方法分为实验组与对照组各36例。实验组与对照组均进行基础治疗和乌司他丁治疗,而实验组还要进行连续性血液净化治疗。记录治疗后APACHEⅡ评分、住院时间、腹痛消失时间、血清淀粉酶恢复时间,测量患者治疗前和治疗一周后的TLR4、血浆炎症因子及白介素水平。结果:(1)实验组患者治疗后住院时间、血清淀粉酶恢复时间、APACHEⅡ评分、腹痛消失时间均显著低于或少于对照组患者(P0.05)。(2)相较于治疗前,两组患者治疗一周后的TLR4、CRP、IL-6和TNF-α水平均显著降低(p0.05);实验组患者治疗一周后的TLR4、CRP、IL-6和TNF-α的水平均显著低于对照组(p0.05)。(3)实验组的治愈率(34.29%)和总有效率(74.29%)均显著高于对照组的治愈率(14.29%)和总有效率(48.57%)。结论:连续性血液净化治疗联合乌司他丁对急性胰腺炎患者的抗炎效果相较于单独使用乌司他丁疗效显著,能更好地降低炎症反应,使得TLR4、血浆炎症因子及白介素等炎症因子降低。     

连续性血液净化及肠内营养治疗重症胰腺炎疗效及对炎症因子影响

摘要 目的：探究连续血液净化及肠内营养治疗重症胰腺炎患者的疗效及对患者炎症因子水平的影响。方法：选择2018年1月至2019年12月于我院接受治疗的82例重症胰腺炎患者为研究对象,按照治疗方式不同将其分为研究组(49例)与对照组(33例)。对照组患者仅接受连续血液净化,研究组患者在对照组患者基础上加用肠内营养支持,对比两组患者腹痛缓解时间、感染发生率、住院时间、治疗前后血尿素氮(Blood urea nitrogen,BUN)、血肌酐(creatinine,SCr)、C反应蛋白(C-reactive protein,CRP)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子(tumor necrosis factor,TNF-α)、急性生理与慢性健康评分表评分(acute physiology and chronic health evaluationⅡ,APACHE II)评分的变化。结果：研究组患者腹痛缓解时间、感染发生率、住院时间均短于对照组患者(P<0.05)。治疗后,研究组患者APACHE II评分、血清CRP、IL-6、TNF-α、BUN及SCr水平均低于对照组(P<0.05)。结论：连续血液净化联合肠内营养支持对重症胰腺炎患者具有较好的治疗效果,能够显著加快重症胰腺炎患者转归,改善患者肾损伤及机体炎症状态。     

乌司他丁联合连续性血液净化治疗重症急性胰腺炎的疗效及对炎性因子和T细胞亚群的影响

目的:探讨乌司他丁联合连续性血液净化(CBP)治疗重症急性胰腺炎(SAP)的疗效及对炎性因子和T细胞亚群的影响。方法:选取2016年5月～2018年9月期间我院收治的SAP患者117例,根据随机数字表法将患者分为对照组(n=58)和研究组(n=59),对照组在基础对症治疗的基础上给予乌司他丁治疗,研究组在对照组基础上联合CBP治疗,比较两组临床疗效、临床各项指标改善情况、炎性因子以及T细胞亚群。结果:研究组治疗后临床总有效率为66.10%(39/59),高于对照组患者的46.55%(27/58)(P0.05)。研究组住院时间、血淀粉酶恢复正常时间、症状缓解时间均短于对照组(P0.05)。两组患者治疗后血清白介素-6(IL-6)、白介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)水平均较治疗前降低,且研究组低于对照组(P0.05)。两组患者治疗后CD4+、CD4+/CD8+均较治疗前升高,且研究组高于对照组(P0.05),CD8+较治疗前降低,且研究组低于对照组(P0.05)。结论:乌司他丁联合CBP治疗SAP患者,疗效显著,可有效改善患者临床症状,提高机体免疫功能,减轻机体炎症反应。     

醋酸奥曲肽联合乌司他丁对重症急性胰腺炎患者血清ET、MCP-1、TNF-α、IL-6水平及预后的影响

目的:探讨醋酸奥曲肽联合乌司他丁对重症急性胰腺炎血清内皮素(ET)、单核细胞趋化因子蛋白1(MCP-1)、肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)水平及预后的影响。方法:纳入我院2017年1月至2018年9月收治的94例重症急性胰腺炎患者,并依据随机数字表法将其分为对照组47例与观察组47例。对照组患者给予乌司他丁治疗,观察组在对照组基础上结合醋酸奥曲肽治疗,两组疗程均为7～14 d。比较两组治疗的疗效,血淀粉酶和尿淀粉酶恢复正常时间,腹痛缓解时间、肠鸣音恢复时间和腹胀缓解时间,治疗前后血清ET、MCP-1、TNF-α、IL-6水平的变化及预后。结果:观察组治疗总有效率(93.62%)显著高于对照组(72.34%)(P0.05)。观察组血淀粉酶和尿淀粉酶恢复时间、腹痛缓解时间、肠鸣音恢复时间和腹胀缓解时间明显短于对照组(P0.05)。两组治疗后血清ET、MCP-1、TNF-α和IL-6水平均较治疗前降低(P0.05),且观察组以上指标均显著低于对照组(P0.05)。观察组出院时生存率高于对照组,但差异无统计学意义(P0.05)。结论:与单用乌司他丁相比,醋酸奥曲肽联合乌司他丁治疗重症急性胰腺炎患者疗效更好,其可显著降低患者血清ET、MCP-1、TNF-α和IL-6水平。     

乌司他丁治疗急性胰腺炎的临床疗效及对患者血清炎性因子水平的影响

目的:观察乌司他丁在急性胰腺炎临床治疗中的作用效果,分析其临床治疗价值。方法:选取我院治疗的急性胰腺炎患者178例,根据治疗方法不同分为观察组和对照组,每组89例。对照组患者采用奥曲肽治疗,观察组患者采用乌司他丁与奥曲肽联合治疗。观察并比较治疗前后患者APACHEⅡ评分和血清炎症因子变化及临床疗效。结果:入院时,两组血清CRP、IL-1、IL-6、TNF-α、APACHEⅡ评分无统计差异(P0.05),治疗后,组内CRP、IL-1、IL-6、TNF-α、APACHEⅡ评分明显下降(P0.05);观察组CRP、IL-1、IL-6、TNF-α、APACHEⅡ评分显著低于参照组(P0.05);观察组临床总有效率(86.52%)显著高于参照组(71.91%),观察组并发症(12.36%)明显低于参照组(24.72%),P0.05,组间有统计差异。结论:乌司他丁能够显著提升急性胰腺炎的临床疗效,促进患者康复,其作用机制可能与抑制炎症因子表达有关。     

连续性血液净化治疗重症急性胰腺炎的临床疗效分析

目的:探讨连续性血液净化治疗重症急性胰腺炎的临床疗效及安全性。方法:选取60例重症急性胰腺炎患者并随机分为两组,观察组(31例)给予连续血液净化治疗,对照组(29例)给予常规治疗。检测盒比较两组治疗前后C反应蛋白(CRP)、谷丙转氨酶(ALT)、氧合指数(PaO_2/FiO_2)、碳酸氢根离子(HCO_3~-)、血肌酐(Scr)水平、急性生理与慢性健康评分(APACHEⅡ)和多器官功能障碍综合征评分(MODS评分)及治疗期间并发症的发生情况和存活情况。结果:治疗后,两组CRP、ALT、HCO3~-、Scr水平均明显下降,PaO_2/FiO_2水平明显升高(P0.05),且观察组CRP、ALT、HCO_3~-、Scr水平低于对照组,而PaO_2/FiO_2水平高于对照组(P0.05)。治疗后,两组APACHEⅡ和MODS评分均较治疗前有所下降,且观察组分值明显低于对照组(P0.05)。两组治疗期间并发症发生率和存活率比较差异无统计学意义(P0.05)。结论:连续性血液净化治疗对重症急性胰腺炎具有较好的治疗效果,能显著改善多器官功能,降低体内炎症反应,调节水电解质平衡,治疗具有较高的安全性。     

生长抑素联合乌司他丁治疗重症急性胰腺炎临床疗效及安全性分析

目的:探讨生长抑素联合乌司他丁治疗重症急性胰腺炎的临床疗效和安全性。方法:将2013年6月至2015年6月期间云南省第三人民医院消化内科收治的96例重症急性胰腺炎患者,随机将其分成对照组和乌司他丁治疗组,每组各48例患者,对照组除常规治疗外给予生长抑素持续静脉滴注,乌司他丁组给予生长抑素联合乌司他丁联合治疗。治疗结束后,比较两组患者治疗总有效率,血清学相关指标及临床指标改善情况,并发症发生情况。结果:乌司他丁治疗组患者治疗总有效率(79.2%)明显高于对照组患者(64.6%)(P0.05);乌司他丁治疗组患者腹痛缓解时间、胃肠减压时间、中转手术率、住院时间及病死率[(3.2±0.5)d,(7.3±2.2)d,4.2%,(15.8±1.5)d,6.3%]均明显低于对照组患者[(4.9±0.6)d,(11.5±3.1)d,10.4%,(24.7±2.1)d,12.5%](P均0.05);乌司他丁治疗组患者治疗后血清淀粉酶、白细胞、C反应蛋白以及白细胞介素6[(140.2±49.1)U/L,(5.2±1.0)×109/L,(6.3±3.4)mg/L,(24.3±4.2)ng/L]均明显低于对照组患者[(430.6±60.2)U/L,(10.2±2.2)×109/L,(16.3±5.2)mg/L,(40.3±5.9)ng/L](P均0.05);乌司他丁治疗组患者并发症ARDS、急性肾功能衰竭、休克发生率(14.6%,12.5%,25.0%)明显低于对照组患者(35.4%,22.9%,39.6%)(P均0.05)。结论:生长抑素联合乌司他丁治疗重症急性胰腺炎疗效显著,能够明显改善患者的血清及临床指标,减少并发症的发生率,值得临床推广应用。     

Predicting rRNA-, RNA-, and DNA-binding proteins from primary structure with support vector machines

In the post-genome era, the prediction of protein function is one of the most demanding tasks in the study of bioinformatics. Machine learning methods, such as the support vector machines (SVMs), greatly help to improve the classification of protein function. In this work, we integrated SVMs, protein sequence amino acid composition, and associated physicochemical properties into the study of nucleic-acid-binding proteins prediction. We developed the binary classifications for rRNA-, RNA-, DNA-binding proteins that play an important role in the control of many cell processes. Each SVM predicts whether a protein belongs to rRNA-, RNA-, or DNA-binding protein class. Self-consistency and jackknife tests were performed on the protein data sets in which the sequences identity was < 25%. Test results show that the accuracies of rRNA-, RNA-, DNA-binding SVMs predictions are approximately 84%, approximately 78%, approximately 72%, respectively. The predictions were also performed on the ambiguous and negative data set. The results demonstrate that the predicted scores of proteins in the ambiguous data set by RNA- and DNA-binding SVM models were distributed around zero, while most proteins in the negative data set were predicted as negative scores by all three SVMs. The score distributions agree well with the prior knowledge of those proteins and show the effectiveness of sequence associated physicochemical properties in the protein function prediction. The software is available from the author upon request.     

Accurate domain identification with structure‐anchored hidden Markov models,saHMMs

The ever increasing speed of DNA sequencing widens the discrepancy between the number of known gene products, and the knowledge of their function and structure. Proper annotation of protein sequences is therefore crucial if the missing information is to be deduced from sequence‐based similarity comparisons. These comparisons become exceedingly difficult as the pairwise identities drop to very low values. To improve the accuracy of domain identification, we exploit the fact that the three‐dimensional structures of domains are much more conserved than their sequences. Based on structure‐anchored multiple sequence alignments of low identity homologues we constructed 850 structure‐anchored hidden Markov models (saHMMs), each representing one domain family. Since the saHMMs are highly family specific, they can be used to assign a domain to its correct family and clearly distinguish it from domains belonging to other families, even within the same superfamily. This task is not trivial and becomes particularly difficult if the unknown domain is distantly related to the rest of the domain sequences within the family. In a search with full length protein sequences, harbouring at least one domain as defined by the structural classification of proteins database (SCOP), version 1.71, versus the saHMM database based on SCOP version 1.69, we achieve an accuracy of 99.0%. All of the few hits outside the family fall within the correct superfamily. Compared to Pfam_ls HMMs, the saHMMs obtain about 11% higher coverage. A comparison with BLAST and PSI‐BLAST demonstrates that the saHMMs have consistently fewer errors per query at a given coverage. Within our recommended E‐value range, the same is true for a comparison with SUPERFAMILY. Furthermore, we are able to annotate 232 proteins with 530 nonoverlapping domains belonging to 102 different domain families among human proteins labelled “unknown” in the NCBI protein database. Our results demonstrate that the saHMM database represents a versatile and reliable tool for identification of domains in protein sequences. With the aid of saHMMs, homology on the family level can be assigned, even for distantly related sequences. Due to the construction of the saHMMs, the hits they provide are always associated with high quality crystal structures. The saHMM database can be accessed via the FISH server at http://babel.ucmp.umu.se/fish/ . Proteins 2009. © 2008 Wiley‐Liss, Inc.     

Protein misfolding, aggregation, and degradation in disease

Pathologies associated with protein misfolding have been observed in neurodegenerative diseases such as Alzheimer’s disease, metabolic diseases like phenylketonuria, and diseases affecting structural proteins like collagen or keratin. Misfolding of mutant proteins in these and many other diseases may result in premature degradation, formation of toxic aggregates, or incorporation of toxic conformations into structures. We review common traits of these diverse diseases under the unifying view of protein misfolding. The molecular pathogenesis is discussed in the context of protein quality control systems consisting of molecular chaperones and intracellular proteases that assist the folding and supervise the maintenance of the folded structure. Furthermore, genetic and environmental factors that may modify the severity of these diseases are underscored. The present article represents a partly revised and updated version of chapter 1 published earlier in volume 232 of the series Methods in Molecular Biology (Walker, J. M., ed., Humana Press, Totowa, NJ), Protein Misfolding and Disease: Principles and Protocols (Bross, P. & Gregersen, N., eds.), pp. 3–16 (2003).     

Chaperonin 60: a paradoxical,evolutionarily conserved protein family with multiple moonlighting functions

Chaperonin 60 is the prototypic molecular chaperone, an essential protein in eukaryotes and prokaryotes, whose sequence conservation provides an excellent basis for phylogenetic analysis. Escherichia coli chaperonin 60 (GroEL), the prototype of this family of proteins, has an established oligomeric‐structure‐based folding mechanism and a defined population of folding partners. However, there is a growing number of examples of chaperonin 60 proteins whose crystal structures and oligomeric composition are at variance with GroEL, suggesting that additional complexities in the protein‐folding function of this protein should be expected. In addition, many organisms have multiple chaperonin 60 proteins, some of which have lost their protein‐folding ability. It is emerging that this highly conserved protein has evolved a bewildering variety of additional biological functions – known as moonlighting functions – both within the cell and in the extracellular milieu. Indeed, in some organisms, it is these moonlighting functions that have been left after the loss of the protein‐folding activity. This highlights the major paradox in the biology of chaperonin 60. This article reviews the relationship between the folding and non‐folding (moonlighting) activities of the chaperonin 60 family and discusses current knowledge on their molecular evolution focusing on protein domains involved in the non‐folding chaperonin functions in an attempt to understand the emerging biology of this evolutionarily ancient protein family.     

Protein design on computers. Five new proteins: Shpilka, Grendel, Fingerclasp, Leather, and Aida.

What is the current state of the art in protein design? This question was approached in a recent two-week protein design workshop sponsored by EMBO and held at the EMBL in Heidelberg. The goals were to test available design tools and to explore new design strategies. Five novel proteins were designed: Shpilka, a sandwich of two four-stranded β-sheets, a scaffold on which to explore variations in loop topology; Grendel, a four-helical membrane anchor, ready for fusion to water-soluble functional domains; Fingerclasp, a dimer of interdigitating β–β–α units, the simplest variant of the “handshake” structural class; Aida, an antibody binding surface intended to be specific for flavodoxin; Leather—a minimal NAD binding domain, extracted from a larger protein. Each design is available as a set of three-dimensional coordinates, the corresponding amino acid sequence and a set of analytical results. The designs are placed in the public domain for scrutiny, improvement, and possible experimental verification.     

Revealing of Proteins Interacting with Na,K-ATPase

Proteins interacting with 11-type of Na,K-ATPase were revealed in pig kidney outer medulla and duck salt glands using three different methods (immunoprecipitation, protein overlay, and chemical cross-linking). Immunoprecipitation was performed after solubilization of protein homogenate with Triton X-100 so that both membrane and cytosol proteins bound to Na,K-ATPase could be revealed. Two other methods were used to study the interaction of cytosol proteins with purified Na,K-ATPase. The sets of proteins revealed by each method in outer medulla of pig kidney were different. Proteins interacting with Na,K-ATPase that have molecular masses 10, 15, 70, 75, 105, 120, and 190 kD were found using the immunoprecipitation method. The chemical cross-linking method revealed proteins with molecular masses 25, 35, 40, 58, 68-70, and 86-88 kD. The protein overlay method revealed in the same tissue proteins with molecular masses 38, 42, 43, 60, 62, 66, 70, and 94 kD.     

The fibril_one on-line database: mutations,experimental conditions,and trends associated with amyloid fibril formation

The association of amyloid fibril formation with a number of important diseases, and the extensive study of this process in vitro, has resulted in a large literature containing a vast amount of information about the fibril formation process. This includes mutations and experimental conditions that promote or protect against fibril formation. A database (fibril_one) was designed to hold information relating to the formation of fibrils. It was populated by extensive searches of the literature and other databases. A powerful World Wide Web query interface to the database was developed, enabling a simple and effective method to view amyloidogenic mutations associated with specific proteins. The Web interface was used to identify trends in the data. This revealed that mutations promoting fibril formation through altered folding tend to be associated with destabilization of the native fold. In particular, tendencies of mutations to disrupt the native secondary structure and packing in the hydrophobic core were discovered to be significant. Query access to the database is available freely on the World Wide Web at http://www.bioinformatics.leeds.ac.uk/group/online/fibril_one.     

一株高蛋白含量小球藻TX的分离鉴定及其诱变育种

【背景】小球藻由于蛋白含量高、营养丰富,在水产养殖上可直接作为鱼、虾、贝类的优质饵料。【目的】对从养殖环境中分离的小球藻进行诱变,选育生长快、蛋白含量高的突变株,为水产养殖天然饵料生产提供优良藻种资源。【方法】以从养殖环境中筛选的生长相对较快且蛋白含量较高的TX作为出发藻株,对该藻株进行分子鉴定,并对该藻株进行紫外诱变、甲基磺酸乙脂(ethyl methyl sulfonate,EMS)诱变和复合诱变,采用96孔板高通量筛选技术和递进式重复筛选方法选育高生物量、高蛋白突变株。【结果】经18SrRNA基因序列分析,TX鉴定为Chlorella sorokiniana,从540个可能的突变株中筛选到8个遗传稳定且生长较快的突变株,其中H10的总蛋白含量达64.2%,可溶性蛋白含量达0.44g/L,干重达0.72g/L,分别较出发藻株提高3.4%、15.8%和26.2%。【结论】突变株H10蛋白含量高且生长较快,可用于天然饵料生产。     

Design,construction, and characterization of a second‐generation DARPin library with reduced hydrophobicity

Designed ankyrin repeat proteins (DARPins) are well‐established binding molecules based on a highly stable nonantibody scaffold. Building on 13 crystal structures of DARPin‐target complexes and stability measurements of DARPin mutants, we have generated a new DARPin library containing an extended randomized surface. To counteract the enrichment of unspecific hydrophobic binders during selections against difficult targets containing hydrophobic surfaces such as membrane proteins, the frequency of apolar residues at diversified positions was drastically reduced and substituted by an increased number of tyrosines. Ribosome display selections against two human caspases and membrane transporter AcrB yielded highly enriched pools of unique and strong DARPin binders which were mainly monomeric. We noted a prominent enrichment of tryptophan residues during binder selections. A crystal structure of a representative of this library in complex with caspase‐7 visualizes the key roles of both tryptophans and tyrosines in providing target contacts. These aromatic and polar side chains thus substitute the apolar residues valine, leucine, isoleucine, methionine, and phenylalanine of the original DARPins. Our work describes biophysical and structural analyses required to extend existing binder scaffolds and simplifies an existing protocol for the assembly of highly diverse synthetic binder libraries.