PHYSICAL-PROPERTIES OF COLLAGEN-SODIUM DODECYL-SULFATE COMPLEXES
PHYSICAL-PROPERTIES OF COLLAGEN-SODIUM DODECYL-SULFATE COMPLEXES
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DOI：
10.1021/bi00588a042
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发表时间：
1979-01-01
期刊：
影响因子：
2.9
通讯作者：
HUDSON, BG
中科院分区：
文献类型：
--
作者：
FREYTAG, JW;NOELKEN, ME;HUDSON, BG
Sodium dodecyl sulfate (NaDodSO4)-polyacrylamide gel electrophoresis and gel filtration chromatography of protein-NaDodSO4 complexes are frequently used to characterize collagen-like polypeptide components in mixtures from basement membrane extracts. Electrophoresis yields anomalously high apparent MW for collagenous polypeptides when typical globular proteins are used as MW standards. The use of gel filtration chromatography for this purpose is suspect because asymmetric particles, including NaDodSO4-protein complexes, coeluted with native globular proteins of lower Stokes radius, when Sepharose 4B is used. To understand these effects and to improve the characterization of collagenous polypeptides, the secondary structure of NaDodSO4-collagen complexes was investigated using circular dichroism, the NaDodSO4 content was measured, the dependence of electrophoretic mobility on gel concentration apparently, and work was extended on gel filtration using a more porous gel, Sepharose CL-4B. The anomalous behavior of [calf] collagen chains on NaDodSO4-polyacrylamide gel electrophoresis was due largely to treatment of data; the method can be used to determine rather accurate values for the number of residues per polypeptide chain. Reliable molecular weights can apparently be obtained using Sepharose CL-4B. These methods can be applied equally well to collagenous and noncollagenous polypeptides.
