Petros A M, Gyenes M, Patthy L, Llinás M
Department of Chemistry, Carnegie-Mellon University, Pittsburgh, Pennsylvania 15213.
Eur J Biochem. 1988 Jan 4;170(3):549-63. doi: 10.1111/j.1432-1033.1988.tb13734.x.
The aliphatic 1H-NMR spectrum of the kringle 4 domain of human plasminogen has been studied via two-dimensional chemical shift correlated (COSY) and nuclear Overhauser correlated (NOESY) experiments at 300 MHz and 620 MHz. A number of aliphatic proton spin systems have been identified and several definite assignments have been made. This was mainly achieved by comparison of the human kringle 4 spectrum with spectra of the porcine, bovine and chicken homologs and also with that of the kringle 1 from human plasminogen on which we have reported previously. The three valyl and two leucyl residues of human kringle 4 have been assigned. The eleven threonyl spin systems have been identified via a RELAYED-COSY experiment and Thr17 has been assigned. The three alanyl spin systems have been identified and assigned. Six seryl spin systems have been identified and the signals from the seven glycyl residues of human kringle 4 have been located with Gly45 assigned. Furthermore, 24 AMX spin systems have been mapped in the COSY spectrum of human kringle 4 and H alpha-H beta,beta' spin systems of Tyr2, Tyr41, Tyr50, Tyr74, Trp25 and Trp62 have been assigned. From the spectrum of a deglycosylated chicken homolog, the epsilon-methyl singlets of Met28 and Met48 have been assigned. Finally, ligand effects on selected aliphatic resonances were observed which could be analyzed in terms of residues likely to neighbor the kringle lysine-binding site.
通过在300兆赫和620兆赫下进行二维化学位移相关（COSY）和核Overhauser相关（NOESY）实验，研究了人纤溶酶原kringle 4结构域的脂肪族¹H-NMR谱。已鉴定出许多脂肪族质子自旋系统，并进行了一些明确的归属。这主要是通过将人kringle 4谱与猪、牛和鸡同源物的谱以及我们先前报道的人纤溶酶原kringle 1的谱进行比较来实现的。已对人kringle 4的三个缬氨酰和两个亮氨酰残基进行了归属。通过接力COSY实验鉴定出11个苏氨酰自旋系统，并对Thr17进行了归属。已鉴定并归属了三个丙氨酰自旋系统。已鉴定出六个丝氨酰自旋系统，并确定了人kringle 4的七个甘氨酰残基的信号，其中Gly45已被归属。此外，在人kringle 4的COSY谱中绘制了24个AMX自旋系统，并对Tyr2、Tyr41、Tyr50、Tyr74、Trp25和Trp62的Hα-Hβ,β'自旋系统进行了归属。从去糖基化鸡同源物的谱中，已对Met28和Met48的ε-甲基单峰进行了归属。最后，观察到配体对选定脂肪族共振的影响，可根据可能邻近kringle赖氨酸结合位点的残基进行分析。
