Amyloid fibrils of glucagon characterized by high-resolution atomic force microscopy

被引:58
作者
De Jong, Kathy L.
Incledon, Bev
Yip, Christopher M.
DeFelippis, Michael R.
机构
[1] Eli Lilly & Co, Lilly Res Labs, Biopharmaceut Res & Dev, Indianapolis, IN 46285 USA
[2] Eli Lilly Canada, Toronto, ON, Canada
[3] Univ Toronto, Toronto, ON, Canada
关键词
D O I
10.1529/biophysj.105.077438
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Glucagon solutions at pH 2.0 were subjected to mechanical agitation at 37 degrees C in the presence of a hydrophobic surface to explore the details of aggregation and fiber formation. High-resolution intermittent- contact atomic force microscopy performed in solution revealed the presence of aggregates after 0.5 h; however, longer agitation times resulted in the formation of fibrillated structures with varying levels of higher-order assembly. Height, periodicity, and amplitude measurements of these structures allowed the identification of four distinct fiber types. The most elementary fiber form, designated a. lament, self-associates in a specific wound fashion to produce protofibrils composed of two filaments. Subsequent self-assembly of these filaments and protofibrils leads to two well-defined brillar motifs, termed Type I and Type II. Atomic force microscopy imaging of pH 2.8 glucagon solutions not agitated or exposed to elevated temperature revealed the presence of amorphous aggregates before the formation of. brillar structures similar to those seen at pH 2.0. Time-course solution Fourier transform infrared spectroscopy and thioflavin T binding studies suggested that glucagon aggregation and fibril formation were associated with the development of b-sheet structure. The results of these studies are used to describe a possible mechanism for glucagon aggregation and fibrillation that is consistent with a hierarchical assembly model proposed for amyloid fibril formation.
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收藏
页码:1905 / 1914
页数:10
相关论文
共 48 条
[1]   Partially folded intermediates in insulin fibrillation [J].
Ahmad, A ;
Millett, IS ;
Doniach, S ;
Uversky, VN ;
Fink, AL .
BIOCHEMISTRY, 2003, 42 (39) :11404-11416
[2]   An amyloid-forming peptide from the yeast prion Sup35 reveals a dehydrated β-sheet structure for amyloid [J].
Balbirnie, M ;
Grothe, R ;
Eisenberg, DS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2001, 98 (05) :2375-2380
[3]   Direct observation of Aβ amyloid fibril growth and inhibition [J].
Ban, T ;
Hoshino, M ;
Takahashi, S ;
Hamada, D ;
Hasegawa, K ;
Naiki, H ;
Goto, Y .
JOURNAL OF MOLECULAR BIOLOGY, 2004, 344 (03) :757-767
[4]   FORMATION AND STRUCTURE OF GELS AND FIBRILS FROM GLUCAGON [J].
BEAVEN, GH ;
GRATZER, WB ;
DAVIES, HG .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1969, 11 (01) :37-&
[5]   ATOMIC FORCE MICROSCOPE [J].
BINNIG, G ;
QUATE, CF ;
GERBER, C .
PHYSICAL REVIEW LETTERS, 1986, 56 (09) :930-933
[6]   Formation of insulin amyloid fibrils followed by FTIR simultaneously with CD and electron microscopy [J].
Bouchard, M ;
Zurdo, J ;
Nettleton, EJ ;
Dobson, CM ;
Robinson, CV .
PROTEIN SCIENCE, 2000, 9 (10) :1960-1967
[7]   Toward understanding insulin fibrillation [J].
Brange, J ;
Andersen, L ;
Laursen, ED ;
Meyn, G ;
Rasmussen, E .
JOURNAL OF PHARMACEUTICAL SCIENCES, 1997, 86 (05) :517-525
[8]  
BRANGE J, 1992, ACTA PHARM NORDICA, V4, P209
[9]  
BROMER WW, 1983, CHEM CHARACTERISTICS, P1
[10]   PREDICTION OF PROTEIN CONFORMATION [J].
CHOU, PY ;
FASMAN, GD .
BIOCHEMISTRY, 1974, 13 (02) :222-245