阳明大学生化所杨永正的Microarray讲座课件.pptVIP

Microarray (II) 楊永正 (yang@.tw) 陽明大學 生化所 June 11, 2001 Integration of information Mechanism Studies Factors involved = Components Order of events = Pathways Interactions = Circuit Microarrays Oligonucleotide array Synthesized on a chip: e.g. Affymetrix Spot on a solid matrix: e.g. Compugen cDNA array: e.g. Incyte Current Applications Fingerprinting Distinguish states (disease vs normal; classification of species, strains, cell lines, drugs, --- etc.) Mechanism studies Pathways and regulatory circuits = Look for drug lead Dynamic changes among developmental stages --- etc. Inducible gene sets are co-regulated. Why do you want to do clustering? Unrelated sequences of similar function cluster together Life phenomena can be represented in a multi-dimensional space Microarray exp. is the nature’s way to classify genes Collect sections from different angles Image reconstruction Database and cross-experiment comparison Most constitutively expressed genes are not regulated Types of reaction pathways Metabolic pathways: modify small molecular substrates Signal transduction pathways: protein modifications Macromolecular synthesis pathways: replication, transcription, translation Processing pathways: protein glycosylation and transport, RNA processing, internalization of receptor Different pathways have different properties They should be analyzed differently; however, they also share common properties 路徑調控與酵素的特性 開放系統 連續多步反應 連續多步反應的調控與生物恆定(homeostasis) 穩定狀態(steady state)由 反應速率決定 改變反應速率的兩種方式 改變反應物濃度 環境變化造成反應物濃度改變 改變酵素量或活性,使反應物濃度改變 改變反應機制:例如 活化酵素: 回饋作用改變KM, Vmax 加入酵素: 環境變化造成新路徑之活化 Methods to change reaction mechanism (KM and Vmax) allosteric interaction covalent modification 異物空間調控效應(allosteric effect) （allos=other; stereos=solid or space）效應與合作（cooperativity）現象 酵素可以與受質接合而改變結構（homotropic allosteric effect） 酵素可以與非受質接合而改變結構 （heterotropic allosteric effect）：迴饋（feedback）現象 酵素兼具感應器,轉化器與開關的功能 以共價鍵修飾酵素亦可產生回饋作用 e.g. pyruvate DH 調控連續多步反應時,訊號應回饋到哪一步驟? 流量控制(flux-generating)反應 什麼是流量控制反應?