STRUCTURAL AND FUNCTIONAL-CHARACTERIZATION OF FULL-LENGTH HEPARIN-BINDING GROWTH ASSOCIATED MOLECULE
STRUCTURAL AND FUNCTIONAL-CHARACTERIZATION OF FULL-LENGTH HEPARIN-BINDING GROWTH ASSOCIATED MOLECULE
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DOI：
10.1091/mbc.3.1.85
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发表时间：
1992-01-01
影响因子：
3.3
通讯作者：
BURGESS, WH
中科院分区：
文献类型：
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作者：
HAMPTON, BS;MARSHAK, DR;BURGESS, WH
肝素结合生长相关分子（HB-GAM）是从成年牛脑和鸡心中分离纯化的。匀浆缓冲液中加入250 mM NaCl后，HB-GAM的产率提高了5- 10倍，表明HB-GAM可能与组织中不溶性成分形成络合物。脑源性HB-GAM的完整氨基酸序列是通过对完整蛋白质和化学或酶源性片段的自动Edman降解建立的。血浆解吸飞行时间质谱法测定的牛HB-GAM的质量为15 291质量单位，与基于氨基酸序列计算的15 289质量单位比较有利。因此，HB-GAM除了信号肽的切割外，没有发生任何主要的翻译后修饰。这些结果表明，先前对该蛋白的氨基酸序列分析是使用截断的HB-GAM进行的。全长HB-GAM不是Balb/3T3克隆A31、Balb MK、NRK或人脐静脉内皮细胞的有丝分裂原。然而，当HB-GAM作为基质呈献给鸡胚大脑皮质源性神经元时，它确实具有粘附特性和神经突延伸活性。HB-GAM作为可溶性因子时，其神经突伸展活性较低。
Heparin-binding growth-associated molecule (HB-GAM) was purified from adult bovine brain and chicken heart. The yield of HB-GAM is increased by 5- to 10-fold when 250 mM NaCl is added to the homogenization buffer, indicating that HB-GAM may exist as a complex with an insoluble component of the tissue. The complete amino acid sequence of the brain-derived HB-GAM was established by automated Edman degradation of the intact protein and chemically or enzymatically derived fragments. The mass of bovine HB-GAM as determined by plasma desorption time-of-flight mass spectrometry is 15 291 mass units, which compares favorably with the calculated mass of 15 289 based on the amino acid sequence. Therefore, HB-GAM has not undergone any major post-translational modifications other than cleavage of the signal peptide. These results indicate that previous amino acid sequence analysis of this protein was carried out using truncated HB-GAM. Full-length HB-GAM is not a mitogen for Balb/3T3 clone A31, Balb MK, NRK, or human umbilical vein endothelial cells. HB-GAM does, however, have adhesive properties and neurite extension activity for chick embryo cerebral cortical derived neurons when presented to these cells as a substrate. HB-GAM had little neurite extension activity when presented as a soluble factor.