Individual cartilage aggrecan macromolecules and their constituent glycosaminoglycans visualized via atomic force microscopy.
Atomic force microscopy was used in ambient conditions to directly image dense and sparse monolayers of bovine fetal epiphyseal and mature nasal cartilage aggrecan macromolecules adsorbed on mica substrates. Distinct resolution of the non-glycosylated N-terminal region from the glycosaminoglycan (GAG) brush of individual aggrecan monomers was achieved, as well as nanometer-scale resolution of individual GAG chain conformation and spacing. Fetal aggrecan core protein trace length (398+/-57 nm) and end-to-end length (257+/-87 nm) were both larger than that of mature aggrecan (352+/-88 and 226+/-81 nm, respectively). Similarly, fetal aggrecan GAG chain trace length (41+/-7 nm) and end-to-end (32+/-8 nm) length were both larger than that of mature aggrecan GAG (32+/-5 and 26+/-7 nm, respectively). GAG-GAG spacing along the core protein was significantly smaller in fetal compared to mature aggrecan (3.2+/-0.8 and 4.4+/-1.2nm, respectively). Together, these differences between the two aggrecan types were likely responsible for the greater persistence length of the fetal aggrecan (110 nm) compared to mature aggrecan (82 nm) calculated using the worm-like chain model. Measured dimensions and polymer statistical analyses were used in conjunction with the results of Western analyses, chromatographic, and carbohydrate electrophoresis measurements to better understand the dependence of aggrecan structure and properties on its constituent GAG chains.
Biological Engineering Division, Massachusetts Institute of Technology, Cambridge, MA, USA.
Laurel
L
Ng
Alan J
AJ
Grodzinsky
Parth
P
Patwari
John
J
Sandy
Anna
A
Plaas
Christine
C
Ortiz
D053674:Aggrecans; D000818:Animals; D002356:Cartilage / Q000737:chemistry*; D002417:Cattle; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans / Q000737:chemistry* / Q000648:ultrastructure; D007091:Image Processing, Computer-Assisted; D037181:Lectins, C-Type; D046911:Macromolecular Substances; D018625:Microscopy, Atomic Force; D015394:Molecular Structure; D010316:Particle Size; D011509:Proteoglycans / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure
D053674:Aggrecans; D016326:Extracellular Matrix Proteins; D006025:Glycosaminoglycans; D037181:Lectins, C-Type; D046911:Macromolecular Substances; D011509:Proteoglycans