Minh Pham

• this paper below can describe what I did as a graduate student at ECU - Biochemistry Department

doi: 10.1007/s10974-005-9053-2.Epub 2006 Feb 1.. 2006;27(1):45-51.J Muscle Res Cell Motil

Minh Pham 1, Joseph M Chalovich

Affiliations expand

• PMID: 16450054
• DOI: 10.1007/s10974-005-9053-2

Fesselin is an actin binding protein from smooth muscle that nucleates actin polymerization in a Ca(++)-calmodulin dependent manner, bundles actin and inhibits the actin-activated ATPase activity of myosin S1. We now report that fesselin binds to smooth muscle alpha-actinin. Binding was measured by blot overlay, affinity chromatography and sedimentation methods. Binding was moderate with an association constant of 1-4 x 10(7) M(-1) assuming a 1:1 association of fesselin with alpha-actinin. Fesselin binds to the central spectrin domain repeat region of alpha-actinin but not to the CH1-CH2 domain. Fesselin accelerates the polymerization of actin. This activity of /fesselin was attenuated by alpha-actinin. These observations support the role of fesselin in organizing the cytoskeleton.

• designed cloning technique for multiple clones, incorporating point mutations into bacteria and virus
• performed protein purification to isolate target proteins for further analysis
• studied interactions of mutated proteins using Biacore to assess binding affinities and kinetics